Strand-Specific Reverse Transcription PCR for Detection of Replicating SARS-CoV-2

Abstract

We developed an assay that detects minus-strand RNA as a surrogate for actively replicating severe acute respiratory syndrome coronavirus 2. We detected minus-strand RNA in 41 persons with coronavirus disease up to 30 days after symptom onset. This assay might inform clinical decision-making about patient infectiousness.

Keywords: 2019 novel coronavirus disease; COVID-19; PCR; RNA; SARS-CoV-2; coronavirus; coronavirus disease; diagnosis; infection control; minus strand; respiratory infections; screening; severe acute respiratory syndrome coronavirus 2; testing; viruses; zoonoses.

Figure 1

Frequency distribution of days between symptom onset and testing in study on strand-specific real-time reverse transcription PCR for detection of replicating severe acute respiratory syndrome coronavirus 2, California, USA, 2020. Dashed line indicates the median number of days since symptom onset. B) Distribution of standard real-time reverse transcription PCR cycle threshold values by results of strand-specific real-time reverse transcription PCR. Horizontal line indicates median.

Figure 2

Deming regression analysis of C_t values by strand-specific real-time reverse transcription PCR as a function of the C_t values by standard real-time reverse transcription PCR for severe acute respiratory syndrome coronavirus 2. Results of PCR for plus strand (A; y = 0.91x + 3.26) and minus strand (B; y = 0.88x + 17.30). C_t, cycle threshold.