Abstract
Background: Infectious Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) was used in the validation of methods for detection of SARS-CoV-2 on stainless-steel surfaces in the AOAC Research Institute Emergency Response Validation project. Handling infectious virus requires Biosafety Level (BSL)-3 facilities.
Objective: To compare the recovery and detection of infectious and heat-inactivated (HI; 65°C for 30 min) SARS-CoV-2 from stainless steel by the modified US Centers for Disease Control and Prevention (CDC) 2019-Novel Coronavirus (2019-nCoV) Real Time Reverse Transcription Polymerase Chain Reaction (RT-PCR) Diagnostic Panel.
Method: Viral stocks were diluted in viral transport medium (VTM) and deposited onto stainless-steel test areas at 2 × 103 and 2 × 104 genomic copies for low and high, respectively. Test areas were sampled and aliquots of the resulting test solutions analyzed by RT-qPCR according to the CDC method. Results were analyzed by probability of detection (POD) statistics.
Results: The low level, where fractional positive results (25-75%) are expected, yielded PODI = 0.80 (0.58, 0.92) for the infectious virus and PODHI = 0.15 (0.05, 0.36) for the HI virus. The bias, dPODHI = -0.65 (-0.80, -0.35), demonstrated a statistical difference between infectious and HI virus detection. No difference was observed at the high inoculation level.
Conclusions: Despite the statistical difference observed, the use of the HI virus is a viable alternative for matrix extension studies using a method comparison study design. Highlights: The use of HI SARS-CoV-2 can mitigate the need for a BSL-3 facility for matrix extension validation of alternative methods in SARS-CoV-2 studies.
Highlights: The use of HI SARS-CoV-2 can mitigate the need for a BSL-3 facility for matrix extension validation of alternative methods in SARS-CoV-2 studies.
【초록키워드】 SARS-CoV-2, 2019-nCoV, Test, prevention, severe acute respiratory syndrome coronavirus-2, RT-PCR, virus, Novel coronavirus, Probability, polymerase chain reaction, RT-qPCR, validation, VTM, CDC, qPCR, response, disease control, virus detection, Control, Study design, infectious, respiratory, genomic, Infectious virus, Emergency, stainless steel, alternative methods, Chain Reaction, alternative method, transport medium, biosafety level, Transport, Viral transport medium, reaction, viral stocks, medium, chain, no difference, positive result, Real time reverse transcription polymerase chain reaction, center, Highlights, extension, heat, Aliquots, mitigate, heat-inactivated, time, statistical, BSL-3 facility, Result, resulting, analyzed, was used, in viral, demonstrated, expected, Level, Panel, POD, 【제목키워드】 detection, prevention, steel, Control, center,