Predicting infectious SARS-CoV-2 from diagnostic samples
Jared Bullard,MD
Kerry Dust,PhD
Duane Funk,MD
James E Strong,MD, PhD
David Alexander,PhD
Lauren Garnett,BSc
Carl Boodman,MD
Alexander Bello,PhD
Adam Hedley,BSc
Zachary Schiffman,BSc
Kaylie Doan,BSc
Nathalie Bastien,PhD
Yan Li,PhD
Paul G Van Caeseele,MD
Guillaume Poliquin,MD, PhD
Corresponding Author: Jared Bullard, 750 William Avenue, Winnipeg, Manitoba, Canada, R3C 3Y1, Email:jared.bullard@gov.mb.ca, Phone: (204)945-1306 Fax: (204)786-4770
Received 2020 May 7.
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Abstract
Background
RT-PCR has become the primary method to diagnose viral diseases, including SARS-CoV-2. RT-PCR detects RNA, not infectious virus, thus its ability to determine duration of infectivity of patients is limited. Infectivity is a critical determinant in informing public health guidelines/interventions. Our goal was to determine the relationship between E gene SARS-CoV-2 RT-PCR cycle threshold (Ct) values from respiratory samples, symptom onset to test (STT) and infectivity in cell culture.
Methods
In this retrospective cross-sectional study, we took SARS-CoV-2 RT-PCR confirmed positive samples and determined their ability to infect Vero cell lines.
Results
Ninety RT-PCR SARS-CoV-2 positive samples were incubated on Vero cells. Twenty-six samples (28.9%) demonstrated viral growth. Median TCID50/ml was 1780 (282-8511). There was no growth in samples with a Ct > 24 or STT > 8 days. Multivariate logistic regression using positive viral culture as a binary predictor variable, STT and Ct demonstrated an odds ratio for positive viral culture of 0.64 (95% CI 0.49-0.84, p<0.001) for every one unit increase in Ct. Area under the receiver operating characteristic curve for Ct vs. positive culture was OR 0.91 (95% CI 0.85-0.97, p<0.001), with 97% specificity obtained at a Ct of >24.
Conclusions
SARS-CoV-2 Vero cell infectivity was only observed for RT-PCR Ct < 24 and STT < 8 days. Infectivity of patients with Ct >24 and duration of symptoms >8 days may be low. This information can inform public health policy and guide clinical, infection control and occupational health decisions. Further studies of larger size are needed.
Keywords: SARS-COV-2, COVID-19, RT-PCR, infectivity, public health