Second round of an interlaboratory comparison of SARS-CoV2 molecular detection assays used by 45 veterinary diagnostic laboratories in the United States. (September 2022)

• Record Type: Journal Article
• Authors: Deng, Kaiping; Uhlig, Steffen; Goodman, Laura B.; Ip, Hon S.; Killian, Mary Lea; Nemser, Sarah M.; Ulaszek, Jodie; Kiener, Shannon; Kmet, Matthew; Frost, Kirstin; Hettwer, Karina; Colson, Bertrand; Nichani, Kapil; Schlierf, Anja; Tkachenko, Andriy; Mlalazi-Oyinloye, Mothomang; Scott, Andrew; Reddy, Ravinder; Tyson, Gregory H.
• Is Part Of: Journal of veterinary diagnostic investigation. Volume 34:Number 5(2022)
• Page Start: 825
• Page End: 834
• Publication Date: 2022-09
• Subjects: interlaboratory comparison -- COVID-19 -- real-time RT-PCR -- SARS-CoV2; Veterinary medicine -- Diagnosis -- Periodicals; 636.0896075
• DOI: 10.1177/10406387221115702
• Languages: English
• ISSNs: 1040-6387

Abstract

The COVID-19 pandemic presents a continued public health challenge. Veterinary diagnostic laboratories in the United States use RT-rtPCR for animal testing, and many laboratories are certified for testing human samples; hence, ensuring that laboratories have sensitive and specific SARS-CoV2 testing methods is a critical component of the pandemic response. In 2020, the FDA Veterinary Laboratory Investigation and Response Network (Vet-LIRN) led an interlaboratory comparison (ILC1) to help laboratories evaluate their existing RT-rtPCR methods for detecting SARS-CoV2. All participating laboratories were able to detect the viral RNA spiked in buffer and PrimeStore molecular transport medium (MTM). With ILC2, Vet-LIRN extended ILC1 by evaluating analytical sensitivity and specificity of the methods used by participating laboratories to detect 3 SARS-CoV2 variants (B.1; B.1.1.7 [Alpha]; B.1.351 [Beta]) at various copy levels. We analyzed 57 sets of results from 45 laboratories qualitatively and quantitatively according to the principles of ISO 16140-2:2016. More than 95% of analysts detected the SARS-CoV2 RNA in MTM at ≥500 copies for all 3 variants. In addition, for nucleocapsid markers N1 and N2, 81% and 92% of the analysts detected ≤20 copies in the assays, respectively. The analytical specificity of the evaluated methods was >99%. Participating laboratories were able to assess their current method performance, identify possible limitations, and recognize method strengths as part of a continuous learning environment to support the critical need for the reliable diagnosis of COVID-19 in potentially infected animals and humans.