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Figure.  Rapid Antigen Test (RAT) and Viral Culture Results
Rapid Antigen Test (RAT) and Viral Culture Results

A, Three individuals tested negative after day 10 (days 12, 13, and 14) but had missing testing data on one or more of the previous days. These individuals were censored to negative at earliest possible negative day (day 11 for 2; day 12 for 1). B, A total of 161 RATs were performed in the 40 participants starting on day 6. Among these, 90 tests were performed in individuals who reported no symptoms on the day of the RAT, with 61 having positive and 29 having negative results (negative predictive value, 32%). C, Viral culture results were obtained from 17 individuals. Lines connect numbers of symptoms for each of the 17 on days 0 and 6.

Table.  Participant Demographic and Clinical Characteristics
Participant Demographic and Clinical Characteristics
1.
Centers for Disease Control and Prevention. Quarantine and isolation. Centers for Disease Control and Prevention. Updated March 30, 2022. Accessed February 22, 2022. https://www.cdc.gov/coronavirus/2019-ncov/your-health/quarantine-isolation.html
2.
Boucau  J, Marino  C, Regan  J,  et al.  Duration of viable virus shedding in SARS-CoV-2 Omicron variant infection.   medRxiv. Preprint posted online March 2, 2022. doi:10.1101/2022.03.01.22271582 Google Scholar
3.
Pekosz  A, Parvu  V, Li  M,  et al.  Antigen-based testing but not real-time polymerase chain reaction correlates with severe acute respiratory syndrome coronavirus 2 viral culture.   Clin Infect Dis. 2021;73(9):e2861-e2866. doi:10.1093/cid/ciaa1706 PubMedGoogle ScholarCrossref
4.
Korenkov  M, Poopalasingam  N, Madler  M,  et al. Evaluation of a rapid antigen test to detect SARS-CoV-2 infection and identify potentially infectious individuals. J Clin Microbiol. 2021;59(9):e0089621. doi:10.1128/JCM.00896-21Crossref
5.
Landon  E, Bartlett  AH, Marrs  R, Guenette  C, Weber  SG, Mina  MJ. High rates of rapid antigen test positivity after 5 days of isolation for COVID-19. medRxiv. Preprint posted online February 2, 2022. doi:10.1101/2022.02.01.22269931
6.
Lefferts  B, Blake  I, Bruden  D,  et al.  Antigen test positivity after COVID-19 isolation—Yukon-Kuskokwim Delta Region, Alaska, January-February 2022.   MMWR Morb Mortal Wkly Rep. 2022;71(8):293-298. doi:10.15585/mmwr.mm7108a3 PubMedGoogle ScholarCrossref
1 Comment for this article
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Analytical Sensitivity of the FlowFlex COVID Ag Home test
Yashpal Agrawal, MBBS PhD | Rocky Mountain Regional VA Medical Center
It is important to note the TCID50/mL value, of the rapid antigen test used in this study. TCID50 represents the dilution of a virus required to infect 50% of a given cell culture. The TCID50 value can also be considered a proxy for the Analytical Sensitivity or the Limit of Detection of a test. Thus the TCID50 for the FlowFlex Home test used in this study is 2500 TCID50 per mL.

In contrast, the widely used BinaxNow COVID-19 Antigen Card and the BinaxNow COVID-19 Home test have a TCID50 of 141/mL. The BinaxNow COVID-19 Self Test on
the other hand has a TCID50 of 160,000/mL.

The BinaxNow COVID-19 Antigen Card test is used quite commonly in the USA and readers should be aware of the significant differences in TCID50 values between tests. Based on its TCID50 of 141/mL, one would theoretically expect a much higher percentage of Antigen positive patients to have culturable virus.
CONFLICT OF INTEREST: None Reported
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Research Letter
Infectious Diseases
August 3, 2022

Duration of Symptoms and Association With Positive Home Rapid Antigen Test Results After Infection With SARS-CoV-2

Author Affiliations
  • 1Division of Infectious Diseases, Brigham and Women’s Hospital, Boston, Massachusetts
  • 2National Emerging Infectious Diseases Laboratories, Boston, Massachusetts
  • 3Department of Microbiology, Boston University School of Medicine, Boston, Massachusetts
  • 4Bioinformatics Program, Boston University, Boston, Massachusetts
  • 5The Broad Institute of MIT and Harvard, Cambridge, Massachusetts
  • 6Center for Computational and Integrative Biology, Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts
JAMA Netw Open. 2022;5(8):e2225331. doi:10.1001/jamanetworkopen.2022.25331
Introduction

Current US Centers for Disease Control and Prevention COVID-19 guidance for nonimmunocompromised individuals allows ending isolation after 5 days if the individual is asymptomatic or afebrile with improving symptoms.1 Culturable virus, currently the best proxy for transmissibility, is reported after day 5.2 It has been proposed that rapid antigen tests (RATs) might assist in determining isolation periods. However, while RATs correlate with culture positivity during early infection,3,4 there are minimal data after day 5, when persistent RAT positivity has been reported.5,6 We sought to compare rates of RAT positivity, COVID-19 symptoms, and positive viral culture starting day 6 after a COVID-19 diagnosis.

Methods

This cohort study was approved by the Mass General Brigham institutional review board. All participants provided online written informed consent. Starting on day 6, individuals newly testing positive tests for SARS-CoV-2 completed an online demographic survey, daily symptom logs, and RAT self-testing. Day 0 was the day of positive SARS-CoV-2 test or symptom onset, whichever came first. On day-6, anterior nasal and separate oral swabs were collected from a convenience sample of 17 individuals (42.5%) for viral culture. Details of the cohort and methods are in the Supplement. The study followed the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) reporting guideline on cohort studies. A t test was used to compare means, using 1-tailed P < .05 to signify statistical significance; R2 was used to explore linear associations between independent variables age, time since last vaccination, and cycle threshold value from initial polymerase chain reaction tests with the dependent variable day of first negative RAT result.

Results

Between January 5 and February 11, 2022, we enrolled 40 individuals (mean [SD] age, 34 [9.5] years; 23 [57.5%] women and 17 [42.5%] men). Of these, 36 (90.0%) had received a primary vaccine series and first booster dose. Details are shown in the Table. None required hospitalization. In this period, 96% to 99% of sequenced isolates in Boston were Omicron BA.1. Only 10 participants (25.0%) had a negative RAT result on day 6, and all had negative results by day 14 (Figure, A). There were no correlations between day of first negative RAT result and age (R2 = 0), time since last vaccine (R2 = 0.05), or cycle threshold value at diagnosis (R2 = 0.03). The mean (SD) day of first negative RAT result in the 7 never-symptomatic participants vs the 33 ever-symptomatic participants was 8.1 (3.0) vs 9.3 (2.4) (P = .14). Positive RAT results were frequent (61 of 90 tests [68%]) on days 6 to 14 among individuals reporting no symptoms that same day (Figure, B).

Seventeen individuals were tested for viral culture on day 6, 12 of whom also had a positive RAT result. Of the 12, 6 had positive culture results (5 anterior nasal and 1 oral) (Figure, C). None had positive results from both sites. No individuals with a negative day-6 RAT result had positive cultures. Of the 6 individuals with positive cultures, 2 reported improving symptoms and 2 reported unchanged symptoms, whereas 2 never reported symptoms. Seven of the 9 reporting no symptoms on day 6 (78%) had negative culture results.

Discussion

In this cohort study of individuals newly diagnosed with COVID-19, 75% continued to have positive RAT results, while 35% had culturable virus on day 6. Everyone with a negative day-6 RAT result had a negative viral culture. However, only 50% of those with a positive RAT result had culturable virus. Acknowledging the caveats of a small cohort of mostly young, vaccinated, nonhospitalized individuals with a presumed Omicron variant and potential variation in self-sampling techniques and lab-based culture methods, these data suggest that a negative RAT result in individuals with residual symptoms could provide reassurance about ending isolation. However, a universal requirement of a negative RAT result may unduly extend isolation for those who are no longer infectious. Meanwhile, a recommendation to end isolation based solely on the presence of improving symptoms risks releasing culture-positive, potentially infectious individuals prematurely, underscoring the importance of proper mask wearing and avoidance of high-risk transmission venues through day 10.

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Article Information

Accepted for Publication: June 19, 2022.

Published: August 3, 2022. doi:10.1001/jamanetworkopen.2022.25331

Correction: This article was corrected on September 23, 2022, to fix an error in the Figure.

Open Access: This is an open access article distributed under the terms of the CC-BY License. © 2022 Cosimi LA et al. JAMA Network Open.

Corresponding Authors: Lisa A. Cosimi, MD, Brigham and Women’s Hospital, 75 Francis St, Boston, MA 02115 (lcosimi@bwh.harvard.edu); Deborah T. Hung, MD, PhD, Broad Institute, 415 Main St, Cambridge, MA 02142 (dhung@broadinstitute.org).

Author Contributions: Drs Cosimi and Hung had full access to all of the data in the study and take responsibility for the integrity of the data and the accuracy of the data analysis.

Concept and design: Cosimi, Seitz, Connor, Hung.

Acquisition, analysis, or interpretation of data: Cosimi, Kelly, Esposito, Seitz, Turcinovic.

Drafting of the manuscript: Cosimi, Esposito, Connor.

Critical revision of the manuscript for important intellectual content: Cosimi, Kelly, Seitz, Turcinovic, Connor, Hung.

Statistical analysis: Cosimi.

Obtained funding: Hung.

Administrative, technical, or material support: Kelly, Esposito.

Supervision: Cosimi, Connor, Hung.

Conflict of Interest Disclosures: Dr Connor reported receiving personal fees from Cell Signaling Technologies, outside the submitted work. Dr Hung reported being a founder of, consultant to, equity holder in, and inventor of technology licensed to Sherlock Biosciences and serving on the scientific advisory board for Proof Diagnostics, outside the submitted work. No other disclosures were reported.

Funding/Support: Costs of this study were covered by operating funds of the Broad Institute of MIT and Harvard. Support for viral culture efforts was also provided by the Massachusetts Consortium on Pathogen Readiness (Dr Connor) and the China Evergrande Group to support viral culture efforts.

Role of the Funder/Sponsor: The Broad Institute’s Environmental Health and Safety Office informed individuals testing positive for COVID-19 of the opportunity to volunteer and consent for the study. The institute’s Procurement and Shipping Office mailed RATs to enrolled individuals.

Additional Contributions: We thank Jesse Souweine, MBA, Kelley Friedman, BS, Michael Nashed, BS, Stacy Gabriel, PhD, and Nicholas Fitzgerald, BS, from the Broad Insitute, Cambridge, Massachusetts, for logistical and organizational support. We thank Cole Sher-Jan, BS, at Boston University, Boston, Massachusetts, for help with rtPCR assays and sample logistics. None received compensation for this work outside of the their usual salaries.

References
1.
Centers for Disease Control and Prevention. Quarantine and isolation. Centers for Disease Control and Prevention. Updated March 30, 2022. Accessed February 22, 2022. https://www.cdc.gov/coronavirus/2019-ncov/your-health/quarantine-isolation.html
2.
Boucau  J, Marino  C, Regan  J,  et al.  Duration of viable virus shedding in SARS-CoV-2 Omicron variant infection.   medRxiv. Preprint posted online March 2, 2022. doi:10.1101/2022.03.01.22271582 Google Scholar
3.
Pekosz  A, Parvu  V, Li  M,  et al.  Antigen-based testing but not real-time polymerase chain reaction correlates with severe acute respiratory syndrome coronavirus 2 viral culture.   Clin Infect Dis. 2021;73(9):e2861-e2866. doi:10.1093/cid/ciaa1706 PubMedGoogle ScholarCrossref
4.
Korenkov  M, Poopalasingam  N, Madler  M,  et al. Evaluation of a rapid antigen test to detect SARS-CoV-2 infection and identify potentially infectious individuals. J Clin Microbiol. 2021;59(9):e0089621. doi:10.1128/JCM.00896-21Crossref
5.
Landon  E, Bartlett  AH, Marrs  R, Guenette  C, Weber  SG, Mina  MJ. High rates of rapid antigen test positivity after 5 days of isolation for COVID-19. medRxiv. Preprint posted online February 2, 2022. doi:10.1101/2022.02.01.22269931
6.
Lefferts  B, Blake  I, Bruden  D,  et al.  Antigen test positivity after COVID-19 isolation—Yukon-Kuskokwim Delta Region, Alaska, January-February 2022.   MMWR Morb Mortal Wkly Rep. 2022;71(8):293-298. doi:10.15585/mmwr.mm7108a3 PubMedGoogle ScholarCrossref
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