Comparison of seven commercial SARS-CoV-2 nucleic acid detection reagents with pseudovirus as quality control material

Yan, Ying; Chang, Le; Luo, Wenxin; Liu, Junyi; Guo, Fei; Wang, Lunan

The ongoing pandemic of COVID-19 threats the whole world, which catalyzes a variety of SARS-CoV-2 nucleic acid test (NAT) kits. To monitor test quality and evaluate NAT kits, quality control materials that best simulate real clinical samples are needed in this study, the performance of SARS-CoV-2 cell culture supernatant, PCDH-based pseudovirus, and MS2-based pseudovirus as quality control materials was compared. As a result, PCDH-based pseudovirus was more similar in characteristics to SARS-CoV-2 particle, and was more suitable for evaluating SARS-CoV-2 NAT kits than MS2-based pseudovirus. Proper detection using sensitive and precise NAT kits is essential to guarantee diagnosis. Thus, limit of detection, precision, anti-inference ability and cross-reactivity of NAT kits from PerkinElmer, Wantai, Shanghai Kehua Bio-Engineering (KHB), Sansure, Da An, BioGerm, and Applied Biological Technologies (ABT) were compared using PCDH-based pseudovirus. For the seven kits evaluated, N gene was more sensitive than ORF1ab gene in most kits, while E gene was most sensitive among the three genes in KHB and ABT. PerkinElmer got the lowest LoD for N gene at 11.61 copies/mL, and 34.66 copies/mL for ORF1ab gene. All the kits showed good precision with CV values less than 5%, as well as acceptable anti-interference ability of 2 mg/L human genomic DNA. No cross-reactivity was observed with other respiratory viruses.