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

Kaiping Deng·Steffen Uhlig·Laura B. Goodman·Hon S. Ip·Mary Lea Killiam·Sarah Nemser·Jodie Ulaszek·Shannon Kiener·Matthew Kmet·Kirstin Frost·Karina Hettwer·Bertrand Colson·Kapil Nichani·Anja Schlierf·Andriy Tkachenko·Mothomang Mlalazi Oyinloye·Scott Andrew·Ravinder Reddy·Gregory H. Tyson
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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.

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Kaiping Deng, Steffen Uhlig, Laura B. Goodman, Hon S. Ip, Mary Lea Killiam, Sarah Nemser, Jodie Ulaszek, Shannon Kiener, Matthew Kmet, Kirstin Frost, Karina Hettwer, Bertrand Colson, Kapil Nichani, Anja Schlierf, Andriy Tkachenko, Mothomang Mlalazi Oyinloye, Scott Andrew, Ravinder Reddy, Gregory H. Tyson. 2022-08-18. Second round of an interlaboratory comparison of SARS-CoV2 molecular detection assays used by 45 veterinary diagnostic laboratories in the United States. https://doi.org/10.1177/10406387221115702

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