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  1. Jan 2022
    1. there are many things I would not have believed about the pandemic pre 2020, but top of them is the fact that Western nations would completely ignore 24 months worth of *actually implemented* policies that were vastly more successful
    1. I've had people mention rising case numbers in Japan and South Korea. But let's really put that rise into perspective. Nations that have early accepted that #COVIDisAirborne simply fair better
    1. Yes, Neoliberal John Snow wins the Internet today H/T @gregggonsalves Reminds me of this….
    1. Update: Switzerland now reports deaths by booster status. Compared to unvaccinated people, the COVID mortality rate is: • 9x lower after full vaccination • 48x lower after a booster [From our post with @maxcroser on death rates by vaccination status: http://ourworldindata.org/covid-deaths-by-vaccination…]
    1. Impact of social distancing OVER a month. 15 vs 406 infections. See below and avoid gatherings
    1. It was always likely that immunity to COVID-19 would wane, and we now know the median time to waning is around 1.5-2 years post infection for most people, sometimes less
    1. That whole opening up going well for Australian inbound tourism then. https://wwwnc.cdc.gov/travel/notices/covid-4/coronavirus-australia
    1. where is the longer term plan for actually making our environments safer to enable us to mix more safely? e.g. cleaner indoor air? better building design for all new builds? We need to take *positive steps* to enable return to normal - not just pretend that Covid has gone away.
    2. This makes it so clear that the release of *all* measures right now (esp masks, esp schools) is only to protect himself & his job. Boris has zero interest in protecting others from getting sick, needing hospital or dying. Or protecting businesses, schools, NHS from disruption.
    1. I took a look at all these NY sequences. I don't think these point mutations S:681H are real. Why? Because they appear all over the Omicron diversity. Some sequences have S:346K, some S:701V, most miss S679K, a few have it. That's the signature of contamination/co-infection.
    1. Kids account for <0.2% of cancer deaths in US Kids account for <0.2% of heart disease deaths in US Kids account for <0.3% of all deaths in the US
    1. This would be an astonishing percentage if @trvrb is right that only 1 in 4 or so cases detected in US.
    1. Pretty big difference US vs UK in this wave. Vaccination differences likely a big part of it.
    1. Just under a million boosters given. 4 confirmed myocarditis cases, 9 under review. All 4 have recovered.
    1. Great thread by @flodebarre on latest genotyping results in France: shows a small increase in potential B.1.640.2 during December in southern France thats now on the decline as Omicron is growing
    2. For those interested first upload of B.1.640.2 onto GISAID was on 4th Nov 2021 from Paris by Roquebert et al, first uploaded Omi was almost 3 weeks later on 22nd Nov 2021 from HK by Alan et al
    3. and as @shay_fleishon pointed out there havent been any new sequences uploaded since before Christmas... this virus has had a decent chance to cause trouble but never really materialised (as far as we can tell at least...)
    4. Lots of chat about B.1.640.2 in the last few days - just a few points to keep in mind: - B.1.640.2 actually predates Omicron - in all that time there are exactly... 20 sequences (compared to the >120k Omis in less time) Def not one worth worrying about too much at the mo...
    1. NL is slightly behind the UK re #omicron and based lockdown on a.o. this model (source @MarionKoopmans). Despite uncertainty the ‘continue as is’ effect on ICU beds occupied (red) is chilling. Green model is with lockdown after and blue is before Christmas. Decisiveness matters!
    1. This is much better. #COVIDisAirborne https://who.int/news-room/questions-and-answers/item/coronavirus-disease-covid-19-how-is-it-transmitted
    1. Interesting. From this afternoon in @nytimes: https://nytimes.com/2022/01/01/us/omicron-covid-holidays-surge-testing.html
    1. Much less loss of smell and taste with Omicron, but more sore throat than with Delta https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/1046623/Technical-Briefing-34-14January2022.pdf… @UKHSA symptom data from ~175,000 Omicron and ~88,000 Delta cases
    1. Maryland - just awful to watch what's unfolding. Now at *1714* hospitalizations 130 in 24 hrs. 16.5% test positivity. Some counties have acted but no statewide policy! No measures to slow the spread. https://coronavirus.maryland.gov
    1. BREAKING: U.S. reports 400,000 new coronavirus cases, the biggest one-day increase on record, with some states yet to report
    1. 544 children with #COVID19 were admitted to U.S. hospitals yesterday— this shattered the previous single-day record that was set 2 days ago (421) source: HHS (https://healthdata.gov/Hospital/COVID-19-Reported-Patient-Impact-and-Hospital-Capa/g62h-syeh…)
    1. #COVID Update: -339,853 Test Results Reported -76,555 Positives -22.53% Positive -7,919 Hospitalizations (+546) -80 new deaths reported by healthcare facilities through HERDS
    1. meet @michaelmina_lab for detailed info on what LFTs are for... https://twitter.com/michaelmina_lab/status/1477831287482241032?s=20
    1. There are sone issues with data from Wales and Northern Ireland covering the holiday weekend, but still, wow. U.K. breaches the 200k daily cases mark for the first time.
    1. How many more days until Rhode Island's cases line exceeds the word "exceeds" in the text header?
    1. Our 6 year old is fully vaccinated He has been in in-person daycare & then elementary school since June 2020 Both daycare & school have required a mask; he wears a well-fitting, comfy 2 layer mask with a KN95 filter as a middle layer—he has had no problem wearing one
    2. Thank you @j_g_allen for continuing to advocate for childhood vaccination & for sharing evidence on masks Yesterday, the U.S. saw a record number of COVID-19 pediatric hospital admissions, almost 1,000 Unvaxxed kids are 10 X more likely to be hospitalized than vaxxed kids 1/2
    1. Good news: admissions for flu not quite as tiny as last year, but close. https://gov.uk/government/statistics/national-flu-and-covid-19-surveillance-reports-2021-to-2022-season…
    1. 3. The geographic breakdown shows some variation, but not tons. The middle of the country has more Delta than anywhere else. The @CDCgov Nowcast can be found here: https://covid.cdc.gov/covid-data-tracker/#variant-proportions…
    2. 2. The @CDCgov Nowcast suggest that 98% of what's out there right now is #Omicron. Less than 2% is Delta. Nothing else currently being spotted.
    3. 1. #Omicron's takeover was stunningly rapid and is now nearly complete, at least in the U.S. The latest "Nowcast" from @CDCgov (which uses recent data to model what's happen now) suggests most of what is circulating here now is omicron.
    1. 5/ PS. As indicated in the chart, even with a booster, vaccines are less effective against Omicron than against Delta. This makes N95 or KN95 masks that much more important.
    2. 4/ Notably, staff seem to be okay. They were required to wear N95s (but students could ware any ole mask). It will be interesting to see if this holds.
    3. 3/ The takeaway should be to get your booster if you qualify (eg, based on when you got your last shot). Also, wear N95s or KN95s. I wish my kids’ middle & high schools had required at least KN95s. Rumor is that a third of the high school is out w/ positive Covid.
    4. 2/ This chart speaks to infection, not hospitalization. Thus, those who were double vaxxed early on (but are not yet boosted) may still have some protection against hospitalization—just not against infection, & it’s unclear how much that protection against hospitalization is.
    5. This chart says it all. If u got double vaccinated early on & are not yet boosted, ur effectively unvaccinated against Omicron infection. Thus, many adults & teens are effectively unvaccinated against infection. Kids got vaccinated recently & thus may be in a safer position. 1/Quote Tweet
    1. A week later and NY State data still showing an extremely high hospitalization VE in the face of Omicron.
    1. Following evidence is that peak infectivity with Omicron comes 3-6 days after symptoms emerge (https://niid.go.jp/niid/en/2019-ncov-e/10884-covid19-66-en.html…), the isolation period is cut to 5 days. How is this 'following the science'
    1. Given ~680k cases per day, this would in turn suggest 0.8% or 1% of the US being infected with SARS-CoV-2 every day. This would translate to perhaps 5% or 10% of individuals currently infected with SARS-CoV-2 in the US. 15/15
    2. I don't have a good sense of how well testing infrastructure held up in London and how this compares to the US, but in general, this would suggest to me to be using something closer to 1 in 4 or 1 in 5 for reporting rate in the US rather than the 1 in 10 I've seen floated. 14/15
    3. This fits with case reports suggesting a large fraction of symptomatic infections for Omicron (https://eurosurveillance.org/content/10.2807/1560-7917.ES.2021.26.50.2101147…, https://medrxiv.org/content/10.1101/2021.12.22.21268021v2…). 13/15
    4. This suggests to me that despite severe outcomes being more rare with Omicron and despite a huge surge in cases that reporting rate for Omicron in London remained fairly stable and did not differ hugely from Delta. 12/15
    5. Importantly if we look at this ratio in Dec during which time Omicron became predominant and case loads increased dramatically, we see that both prevalence and cases increased in tandem and the ~35X ratio was largely maintained. 11/15
    6. If we assume the average infection tests positive for ~10-days (based on @stephenkissler et al https://nejm.org/doi/full/10.1056/NEJMc2102507…), we get an estimated reporting rate of 10/35 = ~29% or very roughly detecting 1 in 3.5 infections as a case. 10/15
    7. We can plot the ratio of prevalence to cases with this 0-day lag to arrive at the following picture through time, where there's some variation, but a ~35X ratio of daily prevalence to daily cases is decently consistent. 9/15
    8. We can compare timeseries of prevalence to cases since Oct 2020. Here we observe a 0-day compatible lag between specimen collection date for cases and prevalence. 8/15
    9. In late December, London had ~26k daily confirmed cases, or 0.3% of the population being recorded as confirmed cases every day. 7/15
    10. Recent data from late Dec from London had ~9% of individuals positive by PCR for SARS-CoV-2. So roughly 1 in 11 people with infections with detectable virus in London. 6/15
    11. Here, I'll be comparing @ONS survey data to @UKHSA case counts in a dataset compiled by @seedragons and available at https://github.com/seedragons/london_covid… (and supplemented with data through Dec 31 via https://ons.gov.uk/peoplepopulationandcommunity/healthandsocialcare/conditionsanddiseases/datasets/covid19infectionsurveytechnicaldata…). 5/15
    12. However, the single best study I'm aware of for estimating reporting rate is the ongoing @ONS study in the UK that mails swabs to a fraction of households regardless of symptom status (https://ons.gov.uk/peoplepopulationandcommunity/healthandsocialcare/conditionsanddiseases/methodologies/covid19infectionsurveypilotmethodsandfurtherinformation…). 4/15
    13. Historically, I have assumed that around 30% of infections in the US are reported as cases. This number was derived from seroprevalence and modeling estimates from sites like (no longer updated) https://covid19-projections.com. 3/15
    14. However, a large fraction of infections, symptomatic and otherwise, don't end up reported as cases due to lack of testing (either the individual doesn't seek testing or testing is desired but not readily found). 2/15
    15. With Omicron, case counts in the US and many other countries have skyrocketed. The US 7-day average is now ~680k cases per day, or 0.2% of the population recorded as confirmed cases each day. 1/15
    1. #COVIDCO2 Quick reference table of rebreathed fraction of air from CO2 measurements, assuming office work levels of oxygen consumption
    2. Here’s the rebreathed fraction of air table from the all-knowing @DavidElfstrom
    1. https://ventilation-mainz.de More than 300 such systems now installed in German schools...if you find this too complicated a simple #corsirosenthalbox filter system will help and can be built in a matter of minutes...https://youtu.be/PtelygpNJQw... failing that windows open + wear masks!
    1. Today at 4! "COVID-19 Vaccines: Science versus Anti-Science" with @PeterHotez. Presented by @huckinstitutes
    1. Our new study: heterogeneous vaccination strategy with inactivated vaccine + mRNA booster elicits strong B and T cell response against the WT and VOC including Omicron @VirusesImmunity
    1. Join the discussion by posting your ideas on problems and solutions in volunteer retention and voluntary sector interactions here: https://padlet.com/groupsandcovid19/b87lztp889ivp9np
    2. Sustaining mutual aid and community support groups: Covid-19 and beyond Free online event, 12.30pm November 3rd
    1. “I want my life back” is a hell of a thing to say much less publish when 5.5 million people have actually lost their actual lives.
    1. Coronavirus variants: Where do they come from? How do we spot them? What do they mean for COVID vaccines, and future of the pandemic?
    1. https://travellingtabby.com/uk-coronavirus-tracker/… The 263 new deaths reported today is the most in a day for almost 8 months. From the 208 new deaths in England, 9 were in individuals below the age of 50, and 1 was a teenager in the 15-19 age range. #Covid19UK #coronavirusuk #DailyCovidUpdate
    1. Russia at 1.04 MILLION excess deaths since March 2020, which is about 240% higher than their reported COVID-19 deaths. This is 1st place worldwide (for countries with data) in absolute excess mortality, 2nd place on per capita terms and 9th on p-score. #poptwitter #epitwitter
    1. Sign of the covid times - front page Scotland edition contrasting with front page London edition
    1. Huge honor to be back @inthebubblepod with @ASlavitt We talked about engaging people with whom we disagree Why disdain for unvaccinated folks is counter-productive And why kindness and understanding (with a side of mandates) will keep our nation in good stead for the long run
    1. .@ASlavitt and @ashishkjha discuss the danger of covering COVID like a political horse race, why he appears on Newsmax so frequently, and how he deals with #COVID skeptics in his own extended family. Listen at http://ow.ly/8jcL50GmwLh
    1. I agree with pretty much all of this @FT article https://ft.com/content/e200156f-2e5a-4165-8aa2-28c24fe3c036
    1. We must protect people with comorbidities from cholera in this 1854 epidemic. I went into medicine – consumption specifically – and public health to protect our most at-risk. Did this tweet alone do enough to protect those most at-risk? Because that's all I'm going to do.
    2. this might be my favourite one so far...
    1. CNN emailed to let me know they are “fact checking” this Tweet & writing an article about it. So I want to be clear, the CDC’s own data says those dying with covid have four or more comorbities & instead of 75% of deaths it’s around 95%. See for yourself. https://cdc.gov/nchs/nvss/vsrr/covid_weekly/index.htm
    2. Here is @ClayTravis showing that he didn't read the article.
    1. Suddenly the CDC admits that 75% of virus deaths were people with 4 or more comorbidities. Seems the “science” now sees the looming 2022 midterm election consequences for lockdown Dems…
    2. Here is @CortesSteve showing that he did not read the article.
    1. BREAKING: CDC Director admits that over 75% of COVID deaths are in people with ‘at least 4 comorbidities.’
    2. here is @jennybethm showing that she did not read the article.
    1. You stupid idiots. You stand in the way of public health. You peabrains do not understand there is no virtue in Omicron-based immunity and mutagenesis. One mutation from Histidine could spell natural disaster you stupid frauds. You are brainless.
    2. T cell immunologist getting very upset at people arguing that high levels of transmission are a good thing
    1. https://travellingtabby.com/scotland-coronavirus-tracker/… Today is the first time in almost three weeks that the positivity rate has been under 20%! Although it is also the first time in about 11 months that we've had over 1,500 people in hospital with the virus. #covid19scotland #DailyCovidUpdate
    1. The US reported 1.4 million COVID cases yesterday and hospitalizations are the highest ever. We’re sailing into this hurricane with virtually no effort to change course because fancy people believe they have a god-given right to their normal lives, no matter how bad things get.
    1. While U.K. daily reported cases fall, COVID deaths (379) up to 28 days after a positive test have sadly hit the highest level since February last year.
    1. The ’with’ or ‘because of’ Covid hospitalization argument is intentionally politicizing data.
    1. Curious if this what @Twitter meant when they talked about their commitment to combat covid disinformation.
    1. Increases in hospitalizations routinely lag behind cases by a week (sometimes more). Deaths have an even longer lag. On 12/30, we had seen a 56% increase in 7-day hospitalization counts, and today we are seeing an 81% increase week-to-week. We have already passed the Sept peak.
    1. If schools are not a source of transmission for COVID-19, why were school board per capita rates of infection 1.77x HIGHER than their surrounding community? SOURCE: Ministry of Education data; Compiled on December 17th, 2021; Calculations are mine.
    1. Good lord! The percent positivity rate in #Ottawa LTC's PCR tests was 83.72% yesterday (while Ottawa large was 50%)!
    1. Teachers on these islands will get FFP2(rightly so).Healthcare workers on other parts of these islands..nah!..Surgical masks/spit guards/not PPE,for working with COVID-positive patients risking other patient’s, our own & our family’s health.”Protect the NHS”⁦@CMO_England
    1. A few tweets on masks for kids (thanks @dgurdasani1). US schools were 3.5 x more likely to have COVID-19 outbreaks if they did not have a mask requirement at the start of school compared with schools that required universal masking on day one. 92/
    1. "Importantly, higher study quality was associated with lower prevalence of all symptoms, except loss of smell & cognitive symptoms" ....as someone who studies cognition I didn't find that as reassuring as possibly intended...
    1. masking is not an "unevidenced intervention" and, at this point, it is outright disinformation to claim so. Sad coming from an academic at a respectable institution
    1. Misinformation on social media is a key challenge to effective and timely public health responses. Existing mitigation measures include flagging misinformation or providing links to correct information but have not yet targeted social processes. Here, we examine whether providing balanced social reference cues in addition to flagging misinformation leads to reductions in sharing behavior. In 3 field experiments (N=817, N=322, and N=278) on Twitter, we show that highlighting which content others within the personal network share and, more importantly, not share combined with misinformation flags significantly and meaningfully reduces the amount of misinformation shared (Study 1-3). We show that this reduction is driven by change in injunctive social norms (Study 2) but not social identity (Study 3). Social reference cues, combined with misinformation flags, are feasible and scalable means to effectively curb sharing misinformation on social media.
    1. Having an incidence of >4000/100.000/14 days (Geneva) is scary & just unbelievable - every day I learn of several friends & colleagues that are positive or were exposed. I wonder how long our household will manage to stay #COVID19 #SARSCoV2 free...
    1. But looking at the wider UK context, Derry is currently looking at 4.5% of everyone *in the entire county* testing positive for COVID in the past week, which is absolutely bonkers. South Wales, the Scottish Central Belt and NW England not too far behind.
    1. The West: "We think you're doing covid pandemic strategy all wrong." Asia: "We don't think of you at all."
    1. Fantastic reassurance for parents. Australian kids it’s time for your jab!!!!Quote TweetEric Topol@EricTopol · 5 JanRemarkably low rate of myocarditis among children mRNA (Pfizer) vaccinated age 5-11 Total of 12 cases among 8.7 million doses given https://cdc.gov/vaccines/acip/meetings/downloads/slides-2022-01-05/02-COVID-Su-508.pdf… Rate of 4/million in boys is <1/10th the rate in teens
    1. BREAKING: U.S. reports 716,714 new coronavirus cases, setting world record for cases in one day
    1. The COVID data coming out of NYC jails is...beyond staggering. Today's report shows a 7-day avg positivity rate of 37%, w/502 ACTIVE INFECTIONS. With a ~5K census, that means that nearly one in ten people in DOC has an ACTIVE infection. Crisis on crisis. https://hhinternet.blob.core.windows.net/uploads/2022/01/CHS-COVID-19-data-snapshot-2020103.pdf
    1. "Eminent" Professor Sunetra Gupta retweets pub landlord. The advancement of "science" 2020-2022...
    1. Warning on death data on https://coronavirus.data.gov.uk NHS England has not reported hospital deaths since 1 January. The backlog will be reported Wednesday. Data are incomplete yesterday, today and tomorrow. Expect a bigger number reported on Wednesday.
    1. NHS Chiefs: 'Hospitals in Crisis' Head Teachers: 'Schools in Crisis' Prime Minister: 'Crisis? What Crisis?' (i.e. you will have to sort it out without any support from us)
    1. A lot of people wear disposable masks.
    2. . I'm guessing it's a good idea to wash/replace them regularly.
    3. Most of it appears to be in this thread actually
    4. So how else could we reduce transmission of an airborne virus if you don’t trust people are savvy enough to get even an ounce of benefit from wearing a mask? Lockdowns and school closures?
    5. Masks are to reduce aerosol inhalation. Nothing to do with washing, touching or snot. As I’m certain you well know.
    6. 4. If a big, snotty sneeze did land within a mask, change the mask (just like one would throw the snotty tissue out if it caught the sneeze). Sanitise hands before putting the fresh mask on.
    7. 1. Wearing a mask will prevent many sneezes as dust and pollens won't get up the nose to cause sneezing. 2. Not all sneezes produce copious snot. 3. If a child is unwell, they should be home, not masked at school.
    8. As you know very effective piblic healtj can be built from very imperfect measures. Masks are one such P.S. if you sneeze, and the mask is dirty, either wipe with a tissue, or swop masks.
    9. CDC has conducted at least 2 large real-life masking studies in public school systems; Yale and partners did an enormous study in Bangladesh. These all support efficacy of masking. Also snot/hand-washing questions are a distraction; disease is predominantly airborne.
    10. German research demonstrates using an FFP2 once a day every week for five weeks works (provided you're not a heavy user). Now that's physics, science, chemistry & using a platform to promote life!
    11. Dear Prof, We’re you aware that this tweet has an anti-mask tone? I hope this was accidental. Would you be able to share proven methods of good face mask hygiene so others can benefit maximally from this potentially highly effective measure against SARS2 inhalation? Thank you
    1. Friday musings - 5 stages of pandemic grief. (I'm sure you'll have ideas too of what to add!)
  2. Dec 2021
    1. Just so I have this clear in my own mind...the Govt has relentlessly pushed the narrative of no need for more restrictions and yet the NHS is building makeshift field hospitals in its car parks
    1. Leeds Hospital to host one of the first 'Nightingale hubs' based in a car park at St James's Hospital. NHS wants space for up to 4k extra beds nationally...but who will staff these beds?
    1. #RobertMalone suspended by #twitter today. Reaction: 1) Great news. He has been spreading harmful #misinformation. (He has NOT contributed to meaningful/constructive scientific debate. His views demonstrably wrong & polarizing.) 2) What took so long? #ScienceUpFirst
    1. 7/ the virus is almost exactly like the flu so there is no need to panic. except it kills lots more people, causes some kind of poorly understood neurological changes, and sometimes lingers for months. but all that’s neither here nor there. basically the same.
    2. 6/ the virus is airborne but it hates perspex. You put a perspex sheet in its path and it self-destructs out of frustration. Doesn’t matter the size of the room or the size of the Perspex.
    3. 5/ related to point 4, the virus hates losses in productivity. Just this past week, a negotiation was held with the virus, and it agreed to hurry and and do what it needs to do quickly so people can get back to work.
    4. 4/ the virus can only infect people that are having a good time and not a boring time. That’s why it spreads at parties and other social gatherings but not at schools or workplaces.
    5. 3/ in restaurants, the virus is temporarily deactivated when patrons are seated at their tables. As soon as they stand to go to the bathroom or bar (for instance) it promptly reactivates.
    6. 2/ for people in a queue, the virus weakens and dies if it must travel perpendicular to the direction of the queue. That’s why people must stand 6 feet apart from the people ahead of and behind them, but not the people next to them in a winding queue. It hates right angles.
    7. 1/ the virus spreads well in indoor settings without proper ventilation (e.g. restaurants), but not when the indoor space was built on the sidewalk or street in New York between August of 2020 and January of 2021.
    8. As a non-virologist, here are some things I have learned about covid that I would not have guessed before the pandemic. They are super counter-intuitive:
    1. Another record day for cases in the U.K yesterday. One of the worst Christmas presents we could have received. Please look after each other as well as you can this festive season.
    1. This is nothing short of scandalous. Unless and until those leading the public health response acknowledge the AIRBORNE nature of the virus and give transmission mitigation advice commensurate with how airborne viruses spread, we will be yo-yoing from wave to wave ad infinitum.
    1. 6.Laboratory of Virology, Division of Intramural Research, National Institute of Allergy 23and Infectious Diseases, National Institute of Health, Hamilton, MT, USA 247.Laboratory of Persistence Viral Diseases, Rocky Mountain Laboratories, National 25Institute of Allergy and Infectious Diseases, National Institute of Health, Hamilton, MT, 26USA 278.Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National 28Institutes of Health, Bethesda, MD, USA 299.National Institute of Dental and Craniofacial Research, National Institutes of Health, 30Bethesda, MD, USA 3110.University of Maryland School of Medicine, Baltimore, MD, USA 3211.Postdoctoral Research Associate Training Program, National Institute of General Medical 33Sciences, National Institutes of Health, Bethesda, MD, USA 3412.R Adams Cowley Shock Trauma Center, Department of Medicine and Program in 35Trauma, University of Maryland School of Medicine, Baltimore, MD, USA 3613.R Adams Cowley Shock Trauma Center, Department of Surgery and Program in Trauma, 37University of Maryland School of Medicine, Baltimore, MD, USA 3814.Department of Medicine, Division of Infectious Disease, University of Maryland School 39of Medicine, Baltimore, MD, USA 4015.Institute of Human Virology, University of Maryland School of Medicine, Baltimore, 41MD, USA 4216.Department of Surgery, Division of Cardiac Surgery, University of Maryland School of 43Medicine, Baltimore, MD, USA 4417.Department of Medicine, Division of Pulmonary and Critical Care Medicine, University 45of Maryland School of Medicine, Baltimore, MD, USA 4618.Hospitalist Department, TidalHealth Peninsula Regional, Salisbury, MD, USA 4719.Division of Critical Care Medicine, Department of Medicine, University of Maryland St. 48Joseph Medical Center, Towson, MD, USA 4920.Medical Virology Section, Laboratory of Infectious Diseases, National Institute of 50Allergy and Infectious Diseases, National Institutes of Health, Bethesda MD 51^See Acknowledgements 52*Corresponding author. Email: chertowd@cc.nih.gov 535455565758596061626364656667COVID-19 is known to cause multi-organ dysfunction1-3 in acute infection, with 68prolonged symptoms experienced by some patients, termed Post-Acute Sequelae of SARS-69CoV-2 (PASC)4-5. However, the burden of infection outside the respiratory tract and time 70to viral clearance is not well characterized, particularly in the brain3,6-14. We performed 71complete autopsies on 44 patients with COVID-19 to map and quantify SARS-CoV-2 72distribution, replication, and cell-type specificity across the human body, including brain, 73from acute infection through over seven months following symptom onset. We show that 74SARS-CoV-2 is widely distributed, even among patients who died with asymptomatic to 75mild COVID-19, and that virus replication is present in multiple pulmonary and 76extrapulmonary tissues early in infection. Further, we detected persistent SARS-CoV-2 77RNA in multiple anatomic sites, including regions throughout the brain, for up to 230 days 78following symptom onset. Despite extensive distribution of SARS-CoV-2 in the body, we 79observed a paucity of inflammation or direct viral cytopathology outside of the lungs. Our 80data prove that SARS-CoV-2 causes systemic infection and can persist in the body for 81months. 82Main text:83 Infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the 84causative agent of coronavirus disease 2019 (COVID-19), has well described pulmonary and 85extrapulmonary manifestations1-3, including multiorgan failure and shock among severe and fatal 86cases. Some survivors experience Post-Acute Sequelae of SARS-CoV-2 (PASC) – also known as 87Long COVID—with cardiovascular, pulmonary, and neurological manifestations with or without 88functional impairment4-5. While autopsy studies of fatal COVID-19 cases support the ability of 89SARS-CoV-2 to infect multiple organs3,7-12, extra-pulmonary organs often lack histopathological 90evidence of direct virally-mediated injury or inflammation10-14. The paradox of extra-pulmonary 91infection without injury or inflammation raises many pathogen- and host-related questions. 92These questions include, but are not limited to: What is the burden of infection within versus 93outside of the respiratory tract? What cell types are infected across extra-pulmonary tissues, and 94do they support SARS-CoV-2 infection and replication? In the absence of cellular injury and 95inflammation in extra-pulmonary tissues, does SARS-CoV-2 persist, and if so, over what 96interval? Does SARS-CoV-2 evolve as it spreads to and persists in different anatomical 97compartments? 98To inform these pathogen-focused questions and to evaluate for the presence or absence 99of associated histopathology in matched tissue specimens, we performed extensive autopsies on 100a diverse population of 44 individuals who died from or with COVID-19 up to 230 days 101following initial symptom onset. Our approach focused on timely, systematic, and 102comprehensive tissue sampling and preservation of adjacent tissue samples for complementary 103analyses. We performed droplet digital polymerase chain reaction (ddPCR) for sensitive 104detection and quantification of SARS-CoV-2 gene targets in all tissue samples collected. To 105elucidate SARS-CoV-2 cell-type specificity and validate ddPCR findings, we performed in situ106hybridization (ISH) broadly across sampled tissues. Immunohistochemistry (IHC) was used to 107further validate cell-type specificity in the brain where controversy remains on the regional 108distribution and cellular tropism of SARS-CoV-2 infection. In all samples where SARS-CoV-2 109RNA was detected by ddPCR, we performed qRT-PCR to detect subgenomic (sg)RNA, an assay 110suggestive of recent virus replication15. We confirmed the presence of replication-competent 111SARS-CoV-2 in extrapulmonary tissues by virus isolation in cell culture. Lastly, in six 112individuals, we measured the diversity and anatomic distribution of intra-individual SARS-CoV-1132 variants using high-throughput, single-genome amplification and sequencing (HT-SGS). 114We categorized autopsy cases of SARS-CoV-2 infection as “early” (n=17), “mid” 115(n=13), or “late” (n=14) by illness day (D) at the time of death, being ≤D14, D15-D30, or ≥D31, 116respectively. We defined persistence as presence of SARS-CoV-2 RNA among late cases. Due to 117the extensive tissue collection, we analyzed and described the results in terms of grouped tissue 118categories as the following: respiratory tract; cardiovascular; lymphoid; gastrointestinal; renal 119and endocrine; reproductive; muscle, skin, adipose, & peripheral nerves; and brain. 120121Autopsy cohort overview 122Between April 26, 2020 and March 2, 2021, we performed autopsies on 44 PCR-123confirmed cases (Extended Data Fig. 1). SARS-CoV-2 seroconversion was detected in 38 of 124these cases (Supplementary Data 1); three early cases (P27, P36, P37) had not seroconverted and 125perimortem plasma was unavailable for the other three cases (P3, P4, P15). Extensive sampling 126of the brain was accomplished in 11 of the 44 cases (Fig. 1). The cohort was 29.5% female with 127a mean age of 59.2 years and was diverse across race and ethnicity (Extended Data Table 1). 12895.5% of patients had at least one comorbidity, with hypertension (54.5%), obesity (52.3%), and 129chronic respiratory disease (34.1%) being most common. Patients presented to the hospital a 130mean of 9.4 days following symptom onset and were hospitalized a mean of 26.4 days. Overall, 131the mean interval from symptom onset to death was 35.2 days and the mean postmortem interval 132was 26.2 hours. 81.8% of patients required intubation with invasive mechanical ventilation, 13322.7% received extracorporeal membrane oxygenation (ECMO) support, and 40.9% required 134renal replacement therapy. Vasopressors, systemic steroids, systemic anticoagulation, and 135antibiotics were commonly administered (Extended Data Table 1). Individual patient-level 136demographic and clinical information can be found in Extended Data Table 2. 137138Widespread infection and persistence 139SARS-CoV-2 RNA was detected in all 44 cases and across 79 of 85 anatomical locations 140and body fluids sampled (Extended Data Fig. 2, Supplementary Data 1). The highest burden of 141SARS-CoV-2 RNA (i.e., >100,000 N gene copies/ng RNA input) was detected in the respiratory 142tract of early cases (Figure 1), but we detected at least 100 N gene copies/ng RNA input from 143every tissue group besides reproductive tissues from multiple individuals among early cases. The 144mean SARS-CoV-2 N gene copies/ng RNA detected from tissues in each grouping among early 145cases are as follows: 9,210.10 across respiratory tissues; 38.75 across cardiovascular tissues; 14630.01 across lymphoid tissues; 24.68 across gastrointestinal tissues; 12.76 across renal and 147endocrine tissues; 0.36 across reproductive tissues; 27.50 across muscle, peripheral nerve, 148adipose, and skin tissues; 57.40 across ocular tissues; and 32.93 across brain tissues (Extended 149Data Table 3). 150With a few exceptions, the overall burden of SARS-CoV-2 RNA decreased by a log or 151more across tissue categories among mid cases, and further decreased among late cases. 152However, several mid and late cases had high levels (≥5 N gene copies/ng RNA input) detected 153among multiple tissues (Extended Data Fig. 2). Further, persistence of low-level SARS-CoV-2 154RNA (0.0004 to <0.5 N gene copies/ng RNA input) was frequently detected across multiple 155tissue categories among all late cases, despite being undetectable in plasma (Extended Data Fig. 1562, Supplementary Data 1). Notably, SARS-CoV-2 RNA was detected in the brains of all six late 157cases and across most locations evaluated in the brain in five of these six, including P42 who 158died at D230 (Fig. 1). 159Overall, SARS-CoV-2 RNA was detected in respiratory tissue of 43/44 cases (97.7%); 160cardiovascular tissue of 35/44 cases (79.5%); lymphoid tissue of 38/44 cases (86.4%); 161gastrointestinal tissue of 32/44 (72.7%); renal and endocrine tissue of 28/44 cases (63.6%); 162reproductive tissue in 17/40 cases (42.5%); muscle, skin, adipose, and peripheral nervous tissue 163in 30/44 cases (68.2%); ocular tissue and humors of 22/28 cases (57.9%); and brain tissue in 16410/11 cases (90.9%) (Extended Data Table 3). 165We additionally detected SARS-CoV-2 sgRNA across all tissue categories, 166predominately among early cases (14/17, 82.4%), as well as in plasma, pleural fluid, and vitreous 167humor (Fig. 1, Extended Data Fig. 2, Supplementary Data 1). sgRNA was also detected in at 168least one tissue of 61.5% of mid cases and 42.9% of late cases, including across three tissue 169categories in a case at D99 (P20). 170We isolated SARS-CoV-2 in cell culture from multiple pulmonary and extrapulmonary 171tissues, including lung, bronchus, sinus turbinate, heart, mediastinal lymph node, small intestine, 172and adrenal gland from early cases up to D7 (P19, P27, P32, P37; Supplementary Data 1). 173174Intra-individual viral variant diversity 175We used HT-SGS to analyze SARS-CoV-2 spike gene variant sequences from a total of 17646 tissues in six individuals. In five individuals from the early group, predominant spike 177sequences were largely identical across tissues. In P27, P19, and P18, no non-synonymous virus 178genetic diversity was detected in pulmonary and extrapulmonary sites despite a high depth of 179single-molecule sampling (Extended Data Fig. 3). Thus, virus populations that were relatively 180homogeneous had disseminated in these individuals without coding changes in spike. However, 181we also noted important patterns of intra-individual virus diversity in several patients from the 182early group. In P27, although all 4,525 inferred spike amino acid sequences were identical, two 183virus haplotypes, each with a single synonymous substitution, were preferentially detected in 184extrapulmonary sites including right and left ventricles and mediastinal LN. In P38, we observed 185clear virus genetic differences between the lung lobes and the brain, with a D80F residue found 186in 31/31 pulmonary but 0/490 brain sequences and a G1219V residue that was restricted to brain 187minor variants. A similar distinction was observed between sequences from dura mater and other 188sites in P36, albeit at very low sampling depth (n = 2 sequences) from dura mater. Overall, these 189findings suggested no need for alterations in receptor utilization to permit extrapulmonary 190dissemination of SARS-CoV-2, while also revealing genetic compartmentalization between 191viruses in the lung lobes and those in extrapulmonary sites, including the brain. 192193ISH reveals SARS-CoV-2 cellular tropism 194We validated our ddPCR results across all tissue categories via ISH for SARS-CoV-2 195spike RNA across selected early, mid, and late cases (Supplementary Data 3). Overall, we 196detected SARS-CoV-2 RNA via ISH in 36 distinct cell types across all sampled organs 197(Extended Data Table 4, Supplementary Data 3). Spike RNA was detected throughout the 198respiratory tract in early cases, as well as within the sinus turbinate, trachea, lungs, from late 199cases (i.e., P33, P20, P42). 200The heart contained spike RNA within myocytes, endothelium, and smooth muscle of 201vessels of both early (P18, P19) and late (P3 & P42) cases. The pericardium demonstrated a 202positive signal for spike RNA within fibroblasts of the stroma. Intimal cells of the aorta were 203additionally found to contain spike RNA. Mononuclear leukocytes within the lymph node, 204spleen, and appendix of an early case (P19) contained spike RNA, as did colonic epithelium (Fig 2052). 206Epithelial cells along the intestinal tract in early cases (P16, P18, P19) contained viral 207RNA, as well as stratified squamous epithelium of the esophagus. Mononuclear leukocytes were 208again visualized with SARS-CoV-2 RNA in lymphoid aggregates and the interstitium of the 209small and large intestine, with infected cells still present in the colon of late cases (P33, P42). 210Kupffer cells, hepatocytes, and bile duct epithelium within the liver were additionally found to 211contain spike RNA. 212Within the kidney, spike RNA could be visualized within parietal epithelium of 213Bowman’s capsule, collecting duct cells, distal tubule cells, and glomerular endothelium. The 214adrenal glands contained spike RNA within endocrine cells. Endocrine follicular cells of the 215thyroid and glandular cells of the pancreas were also positive for spike RNA (Fig. 2). Among 216reproductive organs, spike RNA was visualized within Leydig and Sertoli cells of the testis, 217germ cells within the testicular tubules, endometrial gland epithelium, endometrial stromal cells, 218uterine smooth muscle cells, and stromal cells of the post-menopause ovary (Fig. 2). 219Myocytes within skeletal muscle contained spike RNA in both early (P18) and late (P20) 220cases. In addition to the organ-specific cell type infection of SARS-CoV-2, endothelium, 221muscularis of atrial vessels, and Schwann cells were identified as infected throughout the body, 222and were similarly positive across early and late cases. 223Spike RNA was found in neurons, glia and ependyma, as well as endothelium of vessels 224across all lobes of the brain of early, mid, and late cases. Within the cerebellum specifically, 225neurons, Purkinje cells, and endothelium of vasculature also contained spike protein via IHC 226(Fig. 3). 227228COVID-19 histological findings 229The histopathology findings from our cohort were similar to those reported in other case 230series (Extended Data Fig. 4). All but five cases were considered to have died from COVID-19 231(Extended Data Table 5), and, of these, 37 (94.5%) had either acute pneumonia or diffuse 232alveolar damage at the time of death (Supplementary Data 2). Phases of diffuse alveolar damage 233showed clear temporal associations, with the exudative phase seen mainly within the first three 234weeks of infection and the fibrosing phase not seen until after a month of infection (Extended 235Data Fig. 5). Pulmonary thromboembolic complications, which were also likely related to 236SARS-CoV-2 infection, with or without infarction, were noted in 10 (23%) cases. Another 237finding likely related to SARS-CoV-2 infection included myocardial infiltrates in four cases, 238including one case of significant myocarditis16 (P3). Some of the cases of microscopic ischemia 239appeared to be associated with fibrin-platelet microthrombi, and may therefore be related to 240COVID-19 thrombotic complications. Within the lymph nodes and spleen, we observed 241lymphodepletion and both follicular and paracortical hyperplasia. 242Outside the lungs, histological changes were mainly related to complications of therapy 243or preexisting co-morbidities: mainly obesity, diabetes, and hypertension. Five cases had old 244ischemic myocardial scars and three had coronary artery bypass grafts in place. Given the 245prevalence of diabetes and obesity in our cohort, it was not surprising to find diabetic 246nephropathy (10 cases, 23%) or steatohepatitis (5 cases, 12%). One case was known to have 247chronic hepatitis C with cirrhosis, but the other cases of advanced hepatic fibrosis were likely 248related to fatty liver disease, even if diagnostic features of steatohepatitis were not present. 249Hepatic necrosis (13 cases, 30%) and changes consistent with acute kidney injury (17 cases, 25039%) were likely related to hypoxic-ischemic injury in these very ill patients. 251In the examination of the 11 brains, we found few histopathologic changes, despite the 252evidence of substantial viral burden.Vascular congestion was an unusual finding that had an 253unclear etiology and could be related to the hemodynamic changes incurred with infection. 254Global hypoxic/ischemic change was seen in two cases, one of which was a juvenile (P36) with a 255seizure disorder who was found to be SARS-CoV-2 positive on hospital admission, but who 256likely died of seizure complications unrelated to viral infection. 257258Discussion 259Here we provide the most comprehensive analysis to date of SARS-CoV-2 cellular 260tropism, quantification, and persistence across the body and brain, in a diverse autopsy cohort 261collected throughout the first year of the pandemic in the United States. Our focus on short post-262mortem intervals, comprehensive approach to tissue collection, and preservation techniques –263RNAlater and flash freezing of fresh tissue – allowed us to detect and quantify viral levels with 264high sensitivity by ddPCR and ISH, as well as culture virus, which are notable differences 265compared to other studies. 266We show SARS-CoV-2 disseminates across the human body and brain early in infection 267at high levels, and provide evidence of virus replication at multiple extrapulmonary sites during 268the first week following symptom onset. We detected sgRNA in at least one tissue in over half of 269cases (14/27) beyond D14, suggesting that prolonged viral replication may occur in extra-270pulmonary tissues as late as D99. While others have questioned if extrapulmonary viral presence 271is due to either residual blood within the tissue8,17 or cross-contamination from the lungs during 272tissue procurement8, our data rule out both theories. Only 12 cases had detectable SARS-CoV-2 273RNA in a perimortem plasma sample, and of these only two early cases also had SARS-CoV-2 274sgRNA in the plasma, which occurred at Ct levels higher than nearly all of their tissues with 275sgRNA. Therefore, residual blood contamination cannot account forRNA levels within tissues. 276Furthermore, blood contamination would not account for the SARS-CoV-2 sgRNA or virus 277isolated from tissues. Contamination of additional tissues during procurement, is likewise ruled 278out by ISH demonstrating widespread SARS-CoV-2 cellular tropism across the sampled organs, 279by IHC detecting viral protein in the brain, and by several cases of virus genetic 280compartmentalization in which spike variant sequences that were abundant in extrapulmonary 281tissues were rare or undetected in lung samples. 282Using both ddPCR and sgRNA analysis to inform our selection of tissue for virus 283isolation and ISH staining allow us to describe a number of novel findings. Others6,8-12,17 have 284previously reported SARS-CoV-2 RNA within the heart, lymph node, small intestine, and 285adrenal gland. We demonstrate conclusively that SARS-CoV-2 is capable of infecting and 286replicating within these tissues. Current literature has also reported absent or controversial 287expression of ACE2 and/or TMPRSS2 in several extrapulmonary tissues, such as the colon, 288lymphoid tissues, and ocular tissues, calling into question if these tissues can become infected by 289SARS-CoV-21-3. However, we observed high levels of SARS-CoV-2 RNA and evidence of 290replication within these organs, as well as SARS-CoV-2 RNA via ISH in colonic mucosal 291epithelium and mononuclear leukocytes within the spleen, thoracic cavity lymph nodes, and GI 292lymphoid aggregates. We believe these ISH positive cells represent either infection or 293phagocytized virus in resident macrophages. Further, we isolated virus from a mediastinal lymph 294node and ocular tissue from two early cases (P19, P32). 295Our use of a single-copy sequencing approach for the SARS-CoV-2 spike allowed us to 296demonstrate homogeneous virus populations in many tissues, while also revealing informative 297virus variants in others. Low intra-individual diversity of SARS-CoV-2 sequences has been 298observed frequently in previous studies18-20, and likely relates to the intrinsic mutation rate of the 299virus as well as lack of early immune pressure to drive virus evolution in new infections. It is 300important to note that our HT-SGS approach has both a high accuracy and a high sensitivity for 301minor variants within each sample, making findings of low virus diversity highly reliable21. The 302virus genetic compartmentalization that we observed between pulmonary and extrapulmonary 303sites in several individuals supports independent replication of the virus at these sites, rather than 304spillover from one site to another. Importantly, lack of compartmentalization between these sites 305in other individuals does not rule out independent virus replication, as independently replicating 306populations may share identical sequences if overall diversity is very low. It was also interesting 307to note several cases where brain-derived virus spike sequences showed non-synonymous 308differences relative to sequences from other tissues. These differences may indicate differential 309selective pressure on spike by antiviral antibodies in brain versus other sites, though further 310studies will be needed to confirm this speculation. 311Our results collectively show while that the highest burden of SARS-CoV-2 is in the 312airways and lung, the virus can disseminate early during infection and infect cells throughout the 313entire body, including widely throughout the brain. While others have posited this viral 314dissemination occurs through cell trafficking11 due to a reported failure to culture virus from 315blood3,22, our data support an early viremic phase, which seeds the virus throughout the body 316following pulmonary infection. Recent work by Jacobs et al.22 in which SARS-CoV-2 virions 317were pelleted and imaged from COVID-19 patient plasma, supports this mechanism of viral 318dissemination. Although our cohort is primarily made up of severe cases of COVID-19, two 319early cases had mild respiratory symptoms (P28; fatal pulmonary embolism occurred at home) or 320no symptoms (P36; diagnosed upon hospitalization for ultimately fatal complications of a 321comorbidity), yet still had SARS-CoV-2 RNA widely detected across the body, including brain, 322with detection of sgRNA in multiple compartments. Our findings, therefore, suggest viremia 323leading to body-wide dissemination, including across the blood-brain barrier, and viral 324replication can occur early in COVID-19, even in asymptomatic or mild cases. Further, P36 was 325a juvenile with no evidence of multisystem inflammatory syndrome in children, suggesting 326infected children without severe COVID-19 can also experience systemic infection with SARS-327CoV-2. 328Finally, a major contribution of our work is a greater understanding of the duration and 329locations at which SARS-CoV-2 can persist. While the respiratory tract was the most common 330location in which SARS-CoV-2 RNA tends to linger, ≥50% of late cases also had persistence in 331the myocardium, thoracic cavity lymph nodes, tongue, peripheral nerves, ocular tissue, and in all 332sampled areas of the brain, except the dura mater. Interestingly, despite having much lower 333levels of SARS-CoV-2 in early cases compared to respiratory tissues, we found similar levels 334between pulmonary and the extrapulmonary tissue categories in late cases. This less efficient 335viral clearance in extrapulmonary tissues is perhaps related to a less robust innate and adaptive 336immune response outside the respiratory tract. 337We detected sgRNA in tissue of over 60% of the cohort. While less definitive than viral 338culture23,24, multiple studies have shown that sgRNA levels correlate with acute infection and can 339be detected in respiratory samples of immunocompromised patients experiencing prolonged 340infection24. These data coupled with ISH suggest that SARS-CoV-2 can replicate within tissue 341for over 3 months after infection in some individuals, with RNA failing to clear from multiple 342compartments for up to D230. This persistence of viral RNA and sgRNA may represent infection 343with defective virus, which has been described in persistent infection with measles virus –344another single-strand enveloped RNA virus—in cases of subacute sclerosing panencephalitis25. 345The mechanisms contributing to PASC are still being investigated; however, ongoing 346systemic and local inflammatory responses have been proposed to play a role5. Our data provide 347evidence for delayed viral clearance, but do not support significant inflammation outside of the 348respiratory tract even among patients who died months after symptom onset. 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SARS-CoV-2 genomic and subgenomic 424RNAs in diagnostic samples are not an indicator of active replication. Nat Commun. 42511(1), 6059 (2020). https://doi.org/10.1038/s41467-020-19883-7. 42624.Binnicker, M. J. Can Testing Predict SARS-CoV-2 Infectivity? The Potential for Certain 427Methods To Be Surrogates for Replication-Competent Virus. J Clin Microbiol. 59(11), 428e0046921 (2021). https://doi.org/10.1128/JCM.00469-21. 42925.Sidhu, M. S., et al. Defective measles virus in human subacute sclerosing panencephalitis 430brain. Virology. 202(20), 631-641 (1994). https://doi.org/10.1006/viro.1994.1384. 431432433434435436437Fig. 1 Distribution, quantification, and replication of SARS-Cov-2 across the human body 438and brain. The heat map depicts the highest mean quantification of SARS-CoV-2 RNA (N) via 439ddPCR present within the tissues of eleven COVID-19 autopsy patients who underwent whole 440body and brain sampling. Patients are aligned from shortest to longest duration of illness (DOI) 441prior to death, listed at the bottom of the figure, and grouped into early (≤14 days), mid (15-30 442days), and late (≥31 days) DOI. Tissues are grouped by tissue category beginning with the 443respiratory tract at the top and central nervous system at the bottom. Viral RNA levels range 444from 0.002 to 500,000 N gene copies per ng of RNA input, depicted as a gradient from dark blue 445at the lowest level to dark red at the highest level. Tissues that were also positive for sgRNA via 446real-time RT-PCR are shaded with black vertical bars. L/left, LN/lymph node, NA/not acquired, 447R/right, SC/spinal cord. 448449450451452453454455456457458459460Fig. 2 RNA in situ (RNAscope) detection of SARS-CoV-2 in extrapulmonary tissues. 461SARS-CoV-2 virus is localized to the Golgi and endoplasmic, peri-nuclear in appearance, in the 462following organs and cell types (500 X magnifications): A) Thyroid, demonstrating presence of 463virus within follicular cells. B) Esophagus, demonstrating the presence of virus within the 464stratified squamous epithelium (*), as well as signal in capillaries within the stroma (#). C. 465Spleen, demonstrating the presence of mononuclear lymphoid cells within the white pulp. D) 466Appendix, demonstrating the presence of virus in both colonic epithelium (*) and mononuclear 467lymphoid cells in the stroma (#). E) Adrenal demonstrates virus within endocrine secretory cells 468of the adrenal gland. F) Ovary demonstrates the presence of virus in stromal cells of the ovary in 469a post-menopausal ovary. G) Testis demonstrates the presence of virus in both Sertoli cells (*) 470and maturing germ cells within the seminiferous tubules of the testis (#). H) Endometrium 471demonstrates the presence of virus within endometrial gland epithelium (*) and stromal cells (#), 472in a pre-menopausal endometrial sample. 473474475Fig. 3 SARS-CoV-2 protein expression in human cerebellum. Low magnification 476visualization of no-primary control (A) and primary-added adjacent (B) cerebellar sections 477labeled for SARS-CoV-2 (green) and NeuN (magenta) demonstrate viral-specific protein 478expression within the tissue. The locations of the molecular layer (ML), granular layer (GL), and 479white matter (WM) are indicated in (A) and also correspond to (B). Higher magnification images 480demonstrate cell type-specific infection (C-E). Both NeuN positive neurons (yellow arrows) and 481other unidentified cells (white arrows) are associated with viral protein in the GL (C). Purkinje 482cells adjacent to the ML are infected (D, white arrow). In rare instances, blood vessels adjacent 483to the GL and WM were associated with viral protein (E, white arrow). The scale bars in A is 484also associated with B. All immunofluorescent images were obtained by confocal microscopy. 485486487488489490491492493494495496497498499500501502503504505506Methods: 507Autopsies 508Autopsies were performed and tissues were collected as previously described26 in the National 509Cancer Institute’s Laboratory of Pathology at the National Institutes of Health Clinical Center 510following consent of the legal next of kin. 511512Measurement of IgG and IgM antibodies against Nucleocapsid and Spike protein of SARS-513CoV-2 514Fluid-phase luciferase immunoprecipitation systems (LIPS) assays were used to study IgG and 515IgM antibody response to SARS-CoV-2. For IgG LIPS measurements, Renilla luciferase-516nucleocapsid and Gaussia luciferase-spike protein extracts were employed with protein A/G 517beads (Protein A/G UltraLink Resin, Thermo Fisher Scientific) as the IgG capture reagent as 518previously described with microtiter filter plates27. For IgM measurements, anti-human IgM goat 519agarose beads (Sigma) were substituted as the capture reagent using both the microfilter plate 520and microtube format28. The IgM immunoprecipitation assays performed in 1.5 ml microfuge 521tube format containing 1 l sera or plasma, Renilla luciferase-nucleocapsid (10 million light unit 522input per tube) or Gaussia luciferase-spike protein (40 million light input per tube) and buffer A 523(20 mM Tris, pH 7.5, 150 mM NaCl, 5 mM MgCl2, 0.1% Triton X-100) to a total volume of 100 524l. After mixing, the tubes were incubated at room temp for 1 hour. Next 10 l of the anti-human 525IgM agarose bead suspension was added to each tube for additional 60 minutes and tubes were 526placed on a rotating wheel at 4o C. The samples were then washed by brief centrifugation to 527collect the bead pellet at room temperature 3 times with 1.5 ml Buffer A and once with 1.5 ml of 528PBS. After the final wash, the beads were mixed with coelenterazine substrate (100 l) and light 529units measured in a tube luminometer. Known seronegative and seropositive samples for IgG and 530IgM antibodies against nucleocapsid and spike proteins were used for assigning seropositive cut-531off values and for standardization. 532533 SARS-CoV-2 RNA quantification of tissues and body fluids 534Total RNA was extracted from RNAlater (Invitrogen)-preserved tissues and body fluids 535collected at autopsy using the RNeasy Mini, RNeasy Fibrous Tissue Mini, RNeasy Lipid Tissue 536Mini Kit, and QIAamp Viral RNA Mini Kits (Qiagen) according to the manufacturer’s protocols. 537Upstream tissue processing and subsequent RNA quantification have been previously 538described26. The QX200 AutoDG Droplet Digital PCR System (Bio-Rad) was used to detect and 539quantify SARS-CoV-2 RNA in technical replicates of 5.5 uL RNA for fluids and up to 550 ng 540RNA for tissues as previously described26. Results were then normalized to copies of N1, N2, 541and RP per mL of sample input for fluids and per ng of RNA concentration input for tissues. For 542samples to be considered positive for SARS-CoV-2 N1 or N2 genes, they needed to mean the 543manufacturer’s limit of detection of ≥0.1 copies/μ L and ≥2 positive droplets per well. Over 60 544control autopsy tissues from uninfected patients, representing all organs collected for COVID-19 545autopsy cases, were used to validate the manufacturer’s EUA published LOD for nasopharyngeal 546swabs for tissues (Extended Data Table 8). ddPCR data for P316 as well as a portion of tissues 547from the oral cavity26 have been previously reported. 548549sgRNA analysis of ddPCR positive tissues 550Tissues that tested positive for one or both SARS-CoV-2 N gene targets via ddPCR had RNA 551submitted for sgRNA analysis. Briefly, five μ l RNA was used in a one-step real-time RT-PCR 552assay to sgRNA (forward primer 5’- CGATCTCTTGTAGATCTGTTCTC-3'; reverse primer 5’- 553ATATTGCAGCAGTACGCACACA-3'; probe 5’-FAM-554ACACTAGCCATCCTTACTGCGCTTCG-ZEN-IBHQ-3')29 using the Rotor-Gene probe kit 555(Qiagen) according to instructions of the manufacturer. In each run, standard dilutions of counted 556RNA standards were run in parallel to calculate copy numbers in the samples. The limit of 557detection for this assay was determined to be <40 Cq (Supplemental Data 1) using 40 control 558autopsy tissues from uninfected patients, representing all organs collected for COVID-19 559autopsy cases. 560561Viral isolation from select postmortem tissues 562Select tissues with high viral RNA levels via ddPCR and sgRNA PCR measuring at or below a 56330 Cq underwent virus isolation to prove the presence of infectious virus. Virus isolation was 564performed on tissues by homogenizing the tissue in 1ml DMEM and inoculating Vero E6 cells in 565a 24-well plate with 250 μ l of cleared homogenate and a 1:10 dilution thereof. Plates were 566centrifuged for 30 minutes at 1000 rpm and incubated for 30 minutes at 37°C and 5% CO2. The 567inoculum was then removed and replaced with 500 μ l DMEM containing 2% FBS, 50 U/ml 568penicillin and 50 μg/ml streptomycin. Six days after inoculation, cytopathic effect (CPE) was 569scored. A blind passage of samples where no CPE was present, was performed according to the 570same method. Supernatants from plates with CPE present were analyzed via PCR for SARS-571CoV-2 to rule out other causes of CPE. 572573Virus Sequencing Methods 574Patients with duration of illness ≤7 d (P27, P19) and 8-14 d (P18) with multiple body site 575tissues containing sgRNA levels ≤31 Cq value were selected for high throughput, single-genome 576amplification and sequencing (HT-SGS) as previously described21. Presence of variants of 577SARS-CoV-2 were analyzed within and between tissues. 578579SARS-CoV-2 RNA in situ hybridization 580Chromogenic in situ detection was performed using the manual RNAScope 2.5 HD assay (Cat# 581322310, Advanced Cell Diagnostics, Hayward, CA) with a modified pretreatment protocol.582Briefly, formalin-fixed and paraffin-embedded (FFPE) tissue sections were cut at 7 μm, air dried 583overnight, and baked for 2 hrs at 60ºC. The FFPE tissue sections were deparaffinized, 584dehydrated, and then treated with pretreat 1 for 10 min at room temperature. The slides were 585boiled with pretreatment reagent for 15 min, digested with protease at 40ºC for 10 min, then 586hybridized for 2 hours at 40oC with probe-V-nCov2019-S (Cat# 848561, Advanced Cell 587Diagnostics). In addition, probe-Hs-PPIB (Cat# 313901, Advanced Cell Diagnostics) and probe-588dapB (Cat# 310043, Advanced Cell Diagnostics) were used as a positive and negative control, 589respectively. Subsequent amplification was done according to the original protocol. Detection of 590specific probe binding sites were visualized with RNAScope 2.5 HD Reagent kit-brown 591chromogenic labels (Advanced Cell Diagnostics). The slides were counterstained with 592hematoxylin and cover-slipped. 593594SARS-CoV-2 immunohistochemistry 595FFPE cerebellar sections were deparaffinized, rehydrated and subject to 0.01M Citrate buffer 596antigen retrieval for 20min at 120°C. Slides were incubated in 0.1% TritonX100 in PBS for 59730min, washed extensively with PBS and fresh True Black Plus® solution (1:40, Cat#23014, 598Biotium) applied for 7min. Following PBS wash, blocking serum (5% normal donkey 599serum/0.3M glycine) was applied for 30min. Primary antibodies against SARS-CoV-2 NP1 600(1:250, custom made) and NeuN (1:200, Cat#MAB377, Chemicon) were diluted in blocking 601serum and applied to slides overnight at 4°C. Species-specific secondary conjugates (1:500, 602Cat#A32790 and #A32744, ThermoFisher) were applied for 1hr at RT. Hoescht 33342 applied 603for 10min (1:2000, Cat#H3570, ThermoFisher) labeled nuclei. Slides were cover-slipped with 604Prolong Gold (Cat#P36930, ThermoFisher). 605606Data Availability 607The datasets that support the findings of this study are available in Supplementary Data 1, 2 and 6083. Sequence data described in this manuscript have been deposited (database accession numbers 609XXXX). The bioinformatic pipeline for HT-SGS data analysis has been deposited 610(https://github.com/niaid/UMI-pacbio-pipeline). ISH images from our cohort as well as positive 611and negative controls are available in Supplementary Data 3, which is available at 612https://halo.cancer.gov, Authentication method: NIH, username: halocancernci@gmail.com, 613password: covid19N!H. 614615Methods References: 61626.Huang, N., et al. SARS-CoV-2 infection of the oral cavity and saliva. Nat Med. 27, 892–617903 (2021). https://doi.org/10.1038/s41591-021-01296-8. 61827.Burbelo, P. D., et al. Sensitivity in Detection of Antibodies to Nucleocapsid and Spike 619Proteins of Severe Acute Respiratory Syndrome Coronavirus 2 in Patients With 620Coronavirus Disease 2019. J Infect Dis. 222(2), 206-213 (2020). 621https://doi.org/10.1093/infdis/jiaa273. 62228.Burbelo, P. D., Goldman, R., & Mattson, T. L. A simplified immunoprecipitation method 623for quantitatively measuring antibody responses in clinical sera samples by using 624mammalian-produced Renilla luciferase-antigen fusion proteins. BMC Biotechnol. 5, 22 625(2005). https://doi.org/10.1186/1472-6750-5-22. 62629.Wölfel R., et al. Virological assessment of hospitalized patients with COVID-19. Nature. 627581(7809), 465-469 (2020). https://doi.org/10.1038/s41586-020-2196-x. 628629Acknowledgements: 630This study was funded and supported by the Intramural Research Program of the National 631Institutes of Health, Clinical Center, National Institute of Dental and Craniofacial Research, and 632National Institute of Allergy and Infectious Diseases. 633This research was made possible through the NIH Medical Research Scholars Program, a 634public-private partnership supported jointly by the NIH and contributions to the Foundation for 635the NIH from the Doris Duke Charitable Foundation, Genentech, the American Association for 636Dental Research, and the Colgate-Palmolive Company. 637638NIH COVID-19 Autopsy Consortium 639Daniel S. Chertow1,2, Kevin M. Vannella1,2, Sydney R. Stein1,2, Marcos J. Ramos-Benitez1,2,4, 640Andrew P. Platt1,2, James M. Dickey1,2, Ashley L. Babyak1,2, Luis J. Perez Valencia1,2, Sabrina 641C. Ramelli3, Shelly J. Curran3, Mary E. Richert3, David E. Kleiner5, Stephen M. Hewitt5, Martha 642Quezado5, Willie J. Young5, Sarah P. Young5, Billel Gasmi5, Michelly Sampaio De Melo5, 643Sabina Desar5, Saber Tadros5, Nadia Nasir5, Xueting Jin5, Sharika Rajan5, Esra Dikoglu5, Neval 644Ozkaya5, Kris Ylaya5, Joon-Yong Chung5, Stefania Pittaluga5, Grace Smith5, Elizabeth R. 645Emanuel6, Brian L. Kelsall6, Justin A. Olivera7, Megan Blawas7, Robert A. Star7, Alison 646Grazioli8, Nicole Hays9, Madeleine Purcell9, Shreya Singireddy9, Jocelyn Wu9, Katherine Raja9, 647Ryan Curto9, Jean E. Chung10, Amy J. Borth10, Kimberly A. Bowers10, Anne M. Weichold10, 648Paula A. Minor10, Mir Ahmad N. Moshref10, Emily E. Kelly10, Mohammad M. Sajadi11,12, Kapil 649K. Saharia11,12, Daniel L. Herr13, Thomas M. Scalea14, Douglas Tran15, Ronson J. Madathil15, 650Siamak Dahi15, Kristopher B. Deatrick15, Eric M. Krause16, Joseph Rabin17, Joseph A. Herrold18, 651Ali Tabatabai18, Eric S. Hochberg18, Christopher R. Cornachione18, Andrea R. Levine18, Justin E. 652Richards19, John Elder20, Allen P. Burke20, Michael A. Mazzeffi21, Robert H. Christenson22, 653Zackary A. Chancer23, Mustafa Abdulmahdi24, Sabrina Sopha24, Tyler Goldberg24, Shahabuddin 654Soherwardi25, Yashvir Sangwan26, Michael T. McCurdy27,12, Kristen Sudano27, Diane Blume27, 655Bethany Radin27, Madhat Arnouk27, James W. Eagan Jr28, Robert Palermo29, Anthony D. 656Harris30 657658Affiliations: 6591.Emerging Pathogens Section, Department of Critical Care Medicine, Clinical Center, 660National Institutes of Health, Bethesda, MD, USA 6612.Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, 662Bethesda, MD, USA 6633.Critical Care Medicine Department, Clinical Center, National Institutes of Health, 664Bethesda, MD, USA 6654.Postdoctoral Research Associate Training Program, National Institute of General Medical 666Sciences, National Institutes of Health, Bethesda, MD, USA 6675.Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, National 668Institutes of Health, Bethesda, MD, USA 6696.Mucosal Immunobiology Section, Laboratory of Molecular Immunology, National 670Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, 671USA 6727.Renal Diagnostics and Therapeutics Unit, Kidney Diseases Branch, National Institute of 673Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, 674MD, USA 6758.Kidney Disease Section, Kidney Diseases Branch, National Institute of Diabetes and 676Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA 6779.University of Maryland School of Medicine, Baltimore, MD, USA 67810.University of Maryland Medical Center, Baltimore, MD, USA 67911.Institute of Human Virology, University of Maryland School of Medicine, Baltimore, 680MD, USA 68112.Department of Medicine, Division of Pulmonary and Critical Care Medicine, University 682of Maryland School of Medicine, Baltimore, MD, USA 68313.R Adams Cowley Shock Trauma Center, Department of Medicine and Program in 684Trauma, University of Maryland School of Medicine, Baltimore, MD, USA 68514.Department of Shock Trauma Critical Care, University of Maryland School of Medicine, 686Baltimore, MD, USA 68715.Department of Surgery, Division of Cardiac Surgery, University of Maryland School of 688Medicine, Baltimore, MD, USA 68916.Department of Surgery, Division of Thoracic Surgery, University of Maryland School of 690Medicine, Baltimore, MD, USA 69117.R Adams Cowley Shock Trauma Center, Department of Surgery and Program in Trauma, 692University of Maryland School of Medicine, Baltimore, MD, USA 69318.Department of Medicine, Division of Infectious Disease, University of Maryland School 694of Medicine, Baltimore, MD, USA 69519.Department of Anesthesiology, Division of Critical Care Medicine, University of 696Maryland School of Medicine, Baltimore, MD, USA 69720.Department of Autopsy and Thoracic Pathology, University of Maryland School of 698Medicine, Baltimore, MD, USA 69921.Department of Anesthesiology and Critical Care Medicine, George Washington School 700of Medicine and Health Sciences, Washington, DC USA 70122.Department of Laboratory Science, University of Maryland School of Medicine, 702Baltimore, MD, USA 70323.Department of Anesthesiology, University of Southern California Keck School of 704Medicine, Los Angeles, CA, USA 70524.Critical Care Medicine, University of Maryland Baltimore Washington Medical Center, 706Glen Burnie, MD, USA 70725.Hospitalist Department, TidalHealth Peninsula Regional, Salisbury, MD, USA 70826.Department of Interventional Pulmonology, TidalHealth Peninsula Regional, Salisbury, 709MD, USA 71027.Division of Critical Care Medicine, Department of Medicine, University of Maryland St. 711Joseph Medical Center, Towson, MD, USA 71228.Department of Pathology, University of Maryland, St. Joseph Medical Center, Towson, 713MD, USA 71429.Department of Pathology, Greater Baltimore Medical Center, Townson, MD, USA 71530.Department of Epidemiology and Public Health, University of Maryland School of 716Medicine, Baltimore, MD, USA 717718Author Contributions 719DSC, KMV, SRS, MJRB, ALB, LJPV, AG, DLH, SMH & DEK contributed to the study design 720and protocols for autopsy procurement. APP, JMD, MER, AG, NH, MP, SS, JW, KR, RC, JEC, 721AJB, KAB, AMW, PAM, MANM, EEK, MMS, KKS, DLH, TMS, DT, RJM, SD, KBD, EMK, 722JR, JAH, AT, ESH, CRC, ARL, JER, JE, APB, MAM, RHC, ZAC, MA, SS, TG, SS, YS, MTM, 723KS, DB, BR, MA, JWE Jr, RP, and ADH provided care for, recruited, collected samples from, 724and/or procured medical records for the patients in this study. DEK, SMH, MQ, WJY, SPY, BG, 725MSDM, SD, ST, NN, XJ, SR, ED, NO, KY, JYC, SP, and GS conducted the autopsies and/or 726histological and ISH analysis. SRS, MJRB, APP, JMD, ALB, LJPV, SCR, SJC, ERE, BLK, 727JAO, MB, and RAS assisted with procurement and preservation of autopsy specimens. SRS with 728assistance from SCR and JMD performed RNA extraction, ddPCR, and data analysis. MS, CKY, 729VJM, and EDW performed and analyzed data for sgRNA RT-PCR. CWW and KEP conducted 730IHC on cerebellum. PDB and JIC measured antibody responses to SARS-CoV-2 in perimortem 731plasma samples. SHK, FB, and EAB performed viral sequencing. SRS drafted the manuscript 732with critical input from DSC, KMV, SMH, DEK, SCR, APP, MJRB, EDW, VJM, AG, DLH, 733KKS, MMS MTM, PDB, JIC, CWW, KEP, and SJC. All authors approved the submitted version 734of the manuscript. 735Competing Interests: 736The authors declare no competing or conflict of interest. 737Additional Information: 738Supplementary information is available for this paper. 739Correspondence and requests for materials should be addressed to DSC. 740741742743744745746747748749Extended Data Fig. 1 Autopsy procurement relative to Maryland COVID-19 cases, March 75019th, 2020 to March 9th, 2021. Daily COVID-19 reported cases for Maryland (light blue bars) 751with 7-day average (dark blue line) with timing of autopsies (red arrows). 752753754755756757758759760761762763Extended Data Fig. 2 Distribution, quantification, and replication of SARS-CoV-2 across the 764body and brain over time. The heat map depicts the highest average quantification of SARS-765CoV-2 RNA (N) via ddPCR present within all sampled tissues of 44 autopsy cases. Patients are 766aligned from shortest to longest duration of illness (DOI) prior to death, listed at the bottom of 767the figure, and grouped into early (0-14 d), mid (15-30 d), and late (≥31 d) DOI. Tissues are 768grouped by body system beginning with the respiratory tract at the top and CNS at the bottom. 769Viral RNA levels range from 0.0004 to 500,000 copies per ng of RNA input, depicted as a 770gradient from dark blue at the lowest level to dark red at the highest level. Tissues that were also 771positive for sgRNA via real-time RT-PCR are shaded with black vertical bars. 772773774775776777778779780781782783784785786787Extended Data Figure 3: Analysis of SARS-CoV-2 genetic diversity across body 788compartments in patients. (a) P18, (b) P19, (c) P27, (d) P33, (e) P36, (f) P38. Haplotype 789diagrams (left) show SARS-CoV-2 spike single genome sequences detected in multiple organs. 790Spike NH2-terminal domain (NTD), receptor-binding domain (RBD), and furin cleavage site (F) 791regions are shaded grey, and remaining regions of the spike are shaded white. Ticks with 792different colors indicate mutations relative to the WA-1 reference sequence; green indicates non-793synonymous differences from WA-1 detected in all sequences in the individual; blue indicates 794synonymous mutations detected variably within the individual, and pink indicates non-795synonymous mutations detected variably within the individual. Bar graphs (right) show the 796percentage of all single genome sequences in the sample matching each haplotype. 797798799800801802803804805806807808809810811Extended Data Fig. 4 Representative findings in patients in the COVID-19 cohort. A. Lung, 812Subject P22. Exudative phase diffuse alveolar damage with hyaline membranes and mild 813interstitial inflammation (H&E, 100x). B. Lung, Subject P26. Proliferative phase diffuse alveolar 814damage and sparse inflammation. (H&E, 200x). C. Lung, Subject P22. Organizing thrombus in 815medium sized pulmonary artery. (H&E, 40x). D. Lung, Subject P28. Diffuse pulmonary 816hemorrhage. (H&E, 100x). E. Heart, Subject P3. Active lymphocytic myocarditis with 817cardiomyocyte necrosis. (H&E, 400x). F. Heart, Subject P38. Microscopic focus of bland 818myocardial contraction band necrosis. (H&E, 400x). G. Liver, Subject P41. Steatohepatitis with 819mild steatosis and scattered ballooned hepatocytes. (H&E, 400x), H. Liver, Subject P41. Focal 820bridging fibrosis involving central hepatic veins. (Masson trichrome, 40x). I. Kidney, Subject 821P16. Nodular glomerulosclerosis. (Masson trichrome, 600x). J. Spleen, Subject P16. Preservation 822of white pulp and congestion (H&E, 40x) K. Spleen, Subject P14. Lymphoid depletion of white 823pulp with proteinaceous material and red pulp congestion. (H&E, 100x) L. Spleen, Subject P34. 824Relative preservation of white pulp with extramedullary hematopoiesis (inset) in red pulp (H&E, 825200x) M. Lymph node, Subject P25. Follicular hyperplasia with well-defined follicles. (H&E, ) 826N. Lymph node, Subject P25. Marked plasmacytosis in the medullary cord. (H&E, 400x) O. 827Lymph node, Subject P25. Marked plasmacytosis and sinus histiocytosis. (H&E, 400x) P. Brain, 828Subject P35, Focal subarachnoid and intraparenchymal hemorrhage. (H&E, 40x) Q. Brain, 829Subject P44, Vascular congestion. (H&E, 40x) R. Brain, Subject P43, Intravascular platelet 830aggregates. (anti-CD61 stain, 100x) 831832833834835836837838Extended Data Fig. 5 Temporal association of diffuse alveolar damage in patients dying 839from COVID-19. Number of autopsy cases with stages of diffuse alveolar damage via 840histopathologic analysis by duration of illness. Early time points mainly show the initial 841exudative phase of diffuse alveolar damage, while patients dying after prolonged illness are more 842likely to show organizing or fibrosing stages. 843844845846Extended Data Table 1 Autopsy cohort demographics, comorbidities, and clinical 847intervention summary. (a) Summary of demographics and known comorbidities for autopsy 848cases. (b) Summary of illness course and clinical care for autopsy cases. Data compiled from 849available patient medical records. ECMO/extracorporeal membrane oxygenation. 850851852853854Extended Data Table 2 Individual case demographics and clinical summary. Data obtained 855from available medical records. AF/atrial fibrillation, AVAPS/average volume-assured pressure 856support, BiPAP/bilevel positive airway pressure, CAD/coronary artery disease, CHF/congestive 857heart failure, CKD/chronic kidney disease, CML/chronic myeloid leukemia, COPD/chronic 858obstructive pulmonary disease, DAD/diffuse alveolar damage, DM/diabetes mellitus, DVT/deep 859vein thrombosis, ECMO/extracorporeal membrane oxygenation, ESRD/end-stage renal disease, 860HLD/hyperlipidemia, HTN/hypertension, Hx/historical, ILD/interstitial lung disease, LV/left 861ventricular, MS/multiple sclerosis, PE/pulmonary embolism, PVD/peripheral vascular disease, 862PH/pulmonary hypertension, s/p/status post. 863864865Extended Data Table 3 Summary of SARS-CoV-2 RNA and sgRNA by tissue category over 866time. (a) Summary of the average nucleocapsid gene copies/ng RNA across cases by tissue 867category and duration of illness (days). (b) Summary of the number and percentage of cases with 868SARS-CoV-2 RNA detected via droplet digital (dd)PCR by tissue category for all cases and by 869tissue and duration of illness (days). The number and percentage of tissues positive for ddPCR 870that were additionally positive for subgenomic (sg)RNA PCR is listed in the right most column. 871*A tissue positive via ddPCR was not tested via sgRNA PCR. CNS/central nervous system, 872LN/lymph node. 873874875Extended Data Table 4 SARS-CoV-2 cellular tropism. Summary of cell types that were 876identified as SARS-CoV-2 positive by ISH, and the corresponding anatomic sites in which this 877was observed. 878Cell TypeLocationsBile duct epitheliumLiverChondrocytesBronchial cartilage ringsCollecting duct epitheliumKidneyDistal tubule epitheliumKidneyEndocrine cells of adrenalAdrenal glandEndocrine cells of thyroidThyroidEndotheliumVasculature, allEpendymaBrain Exocrine cells of pancreasPancreasFibroblast-like cellsPericardium, heart, trachea, bronchusGerm cellsTestisGlandular epithelumUterus GliaBrain, all locationsHepatocytesLiverHyaline MembraneLungInterstitial cells of endometriumUterusIntimal cellsAortaKupffer cellsLiverLeydig cellsTestisMononuclear leukocytesLung, spleen, lymph nodes, lymphoid aggregates of GIMucosal epitheliumSmall intestine, colonMucus secreting epithelium, salivary typeSalivary glands, trachea, bronchusMyocytes, CardiacHeartMyocytes, StriatedPsoas muscleMyocytes, SmoothUterus, GINeuronsBrain, all locationsParietal cells Kidney, Bowman's capsulePneumocytes, type I & IILungPurkinje cellCerebellumSchwann cellsNerves, allSertoli cellsTestisStratified epithelium (& basal layer)Trachea, esophagusStromal cellsPericardium, uterus, ovaryVascular smooth muscleArteries, all879Extended Data Table 5 Histopathologic findings of COVID-19 autopsy cases. Summary of 880histopathologic findings across organ system across 44 autopsy cases. Central nervous system 881findings are reported for the 11 cases in which consent for sampling was obtained. 1Includes one 882case in which the COVID lungs were transplanted and data from explanted lungs used in table. 8832Individual lung weights were missing in 4 cases. 3Findings missing on 1 case due to extreme 884autolysis. 4Weight missing on one case. 5Lymph node findings missing in 4 cases 885
    1. That awful VAERS preprint, championed by anti-vaxxers Marjorie Taylor Greene & Robert Malone, which argued the vax is more dangerous than Covid, refuses to die The preprint was coauthored by a member of Rational Ground, an extreme Covid denialist group that harasses scientists
    1. A study in @NatureMedicine indicates that the effectiveness of the Pfizer/BioNTech and Oxford-AstraZeneca vaccines against SARS-CoV-2 infections with symptoms or high viral burden is reduced with the Delta variant compared to the Alpha variant. https://go.nature.com/3vecpoj
    1. 1/ The regular vaccines give great booster responses against Delta and other variants This was first shown by Moderna, with 'original recipe' vaccine booster, compare to Beta booster. https://nature.com/articles/s41591-021-01527-y
    1. Just published @ScienceTM With blunting of the immune response, why it is so important for pregnant women to get a Covid vaccine https://science.org/doi/10.1126/scitranslmed.abm2070… w/ @ScienceVisuals
    1. Politics is derailing a crucial debate over the immunity you get from recovering from #Covid19 https://statnews.com/2021/10/19/politics-is-derailing-a-crucial-debate-over-the-immunity-you-get-from-recovering-from-covid-19/… @levfacher via @statnews “People on the right scream, so people on the left say no. We’re in this horrible, awful feedback loop of vitriol right now.”
    1. There are 2 other impressive conclusions from this study: 1. Comparing vaccine effectiveness of booster vs 'fully vaxxed' as the baseline. Booster ADDS 81-85% protection against symptomatic infection ON TOP of what you already had from your primary (2-dose) vaccination
    1. Comprehensive dossier evidence of effectiveness of third dose Vax. @TVMohandasPai @ARanganathan72 @TimesNow @RShivshankar
    1. A dose of the AstraZeneca COVID-19 vaccine followed by a dose of the Pfizer/BioNTech vaccine confers better protection against SARS-CoV-2 infection than two doses of the Pfizer vaccine, according to a study in healthcare workers published in @Nature. https://go.nature.com/3npW5gO
    1. 'The benefit of a third [booster] dose in reducing transmission is sizeable and increases with vaccine coverage and contact rates among individuals." https://medrxiv.org/content/10.1101/2021.10.25.21265500v1… by @billy_gardner_ and @DiseaseEcology
    1. 3.5% of health and care workers already have long COVID- the highest across all occupations. How can we expect the NHS to manage a huge crisis when our govt is doing very little to support it?
    1. As Omicron reality dawns, consider investing in better masks: "....tight-fitting FFP2 masks provide *75 times* better protection compared to well-fitting surgical masks"
    1. Wow. COVID vaccine misinformation continues to be soooo horrible. This is incredible widespread and ABSOLUTELY made up. (Just like the insanity of implantable chips they continue to claim over and over) These fabrications are so damaging to the health of Americans.
    1. How do we get broad immunity to SARS-CoV-2 that will protect against future variants? 2 studies (are there more?) suggest that vaccination followed by infection gives broader protection than infection followed by vaccination. @florian_krammer @profshanecrotty @GuptaR_lab
    1. EMA recommends approval of BioNTech/Pfizer’s #COVID19vaccine, Comirnaty, for children aged 5 to 11. In this population, the dose of #Comirnaty will be lower than that used in people aged 12 and above. Read the full press release: https://ema.europa.eu/en/news/comirnaty-covid-19-vaccine-ema-recommends-approval-children-aged-5-11
    1. Five quick tweets on the new variant B.1.1.529 Caveat first: data here is *very* preliminary, so everything could change. Nonetheless, better safe than sorry. 1) Based on the data we have, this variant is out-competing others *far* faster than Beta and even Delta did
    1. Around the world, there are now 132 COVID-19 vaccine candidates undergoing clinical trials and 194 candidates in pre-clinical development. Read our latest #COVID19 vaccine race update:
    1. “Participants who received a booster at least 5 months after a second dose of BNT162b2 had 90% lower mortality due to Covid-19 than participants who did not receive a booster.” #covid19
    1. We find that logistic growth of Omicron sequence fraction looks similar between the UK, the US and Germany with roughly 1% of sequenced cases in all three countries being Omicron on Dec 1. 3/10
    1. Well captured by @snolen. Even as vaccine supply becomes more reliable, the uptake challenge across Africa is partly down to “vaccine indifference” rather than hesitancy––there are far more pressing problems across the region.
    1. "There is no evidence currently that COVID-19 vaccines are linked to an increase in sportspeople collapsing or dying due to heart issues such as myocarditis."
    1. This is an important point that needs to get out there. It takes time for the *booster* to work. People will not have invincibility straight after receiving their booster, and should still be cautious.
    1. As well as Tom's new one (B.1.1.529), C.1.2 seems to be spreading in S Africa - C.1.2 was the one with lots of worrying mutations first reported in August... plus cases in S Africa suddenly increasing again in the middle of their summer.
    1. 2 key resources, if you writing about us in Africa right now: Africa-Vaccine Data 2021: https://mediahack.co.za/datastories/coronavirus/africa-covid19/africa-vaccinations/… @mediahackza South Africa Vaccine Deliveries Quarterly, 2021: https://healthjusticeinitiative.org.za/2021/10/13/hji-summary-sheets-vaccine-supplies/#dearflip-df_3476/1/… @HealthJusticeIn Also: @Bhekisisa_MG @Afri_Alliance @PHMSA1 @MSF_access
    1. NEW translation of the Covid-19 Handbook for our friends in Poland https://c19vax.scibeh.org/pl This work was only possible because of the work done by Mateusz Kozak and Beata Kuśmider.
    1. but it is not vaccinated people that are disproportionately filling up ICUs. For any government whose policy is guided by ICU capacity, limiting the transmission possibilities for the unvaccinated is now the point. It is frustrating to see someone continue to ignore this
    1. #Omicron update from Denmark. Omicron cases on the 12th of December adjusted up to 20.5% (+2%). Omicron is now having a large impact on the total number of cases in Denmark. The government has announced a press-meeting Friday, and PM says new measures are needed. 1/4
    1. Current excess deaths estimate per @TheEconomist: 18.2 million with 95% range of 11.4 million to 21.2 million. And up it will go. https://economist.com/graphic-detail/coronavirus-excess-deaths-estimates?fsrc=core-app-economist
    1. No evidence that Omicron is less severe, new @imperialcollege study reports https://ft.com/content/020534b3-5a54-4517-9fd1-167a5db50786
    1. South Africa Deaths (7 day average) are near double levels of a few weeks ago. They will go up further. They are *only* 7.5% of Delta peak, despite higher cases. It should NOT be a surprise that more deaths are being reported after so many cases and hospitalizations.
    1. Exclusive: Hospitals plan for ‘mass casualty’ event with up to one-third of staff sick
    1. positivity rising steeply too - there are definitely many more people we aren't catching
    1. COVID-19 & B.1.1.529: a chance for govt & others to change their mind and their approach? While we are still gathering data on B.1.1.529, the early indicators are very very alarming- likely far more transmissible and more serious than delta + more vaccine escape. (1/10)
    1. vaccine equity has been a disaster, but I do wonder whether the exclusive focus on donations does the US/EU comparison justice. The EU allowed the export of huge numbers of EU produced doses at a time when the US did not (and EU itself was struggling to meet demand).
    1. …this info is more relevant than immune escape. For instance the B.1.351 from ZA last year never really took off. So transmissibility a greater factor for global spread. But we’ll learn more in coming days. Based on inadequate genomic sequencing in US it’s likely here already
    1. maybe this is a good moment to remind people that makers of mRNA vaccines have been extensively prepping for the possibility of new variants. Biontech/Pfizer have given a timeline of 100 days to the delivery of a retooled version of their vaccine
    1. Parts of Germany seem to have potentially introduced requirements that cannot practically be met as testing capacity is proving insufficient - a dangerous moment for rule compliance Nadelöhr Corona-Tests: "Es ist Wahnsinn" via @sz
    1. Following the meeting of the @WHO TAG-VE today, WHO classifies B.1.1.529 as a variant of concern named Omicron. We call for increased surveillance and genetic testing & thank for sharing their work in real time. More information can be found here https://who.int/news/item/26-11-2021-classification-of-omicron-(b.1.1.529)-sars-cov-2-variant-of-concern
    1. German statistics suggesting that 8-9 of 10 infections involve at least one unvaccinated person