Comparison of Seven Commercial Severe Acute Respiratory Syndrome Coronavirus 2 Nucleic Acid Detection Reagents with Pseudovirus as Quality Control Material.

ABSTRACT

The ongoing pandemic of coronavirus disease 2019 threatens the whole world, which catalyzes a variety of severe acute respiratory syndrome coronavirus 2 (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. PCDH-based pseudovirus was found to be more similar in characteristics to SARS-CoV-2 particle, and 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, Beijing Wantai Biological Pharmacy Enterprise Co, Ltd, Shanghai Kehua Bio-Engineering Co, Ltd, Sansure Biotech Inc., Da An Gene Co, Ltd, Shanghai BioGerm Medical Biotechnology Co, Ltd, and Applied Biological Technologies Co, Ltd, were compared using PCDH-based pseudovirus. For the seven kits evaluated, N gene was more sensitive than ORF1ab gene in most kits, whereas E gene was most sensitive among the three genes in Shanghai Kehua Bio-Engineering Co, Ltd, and Applied Biological Technologies Co, Ltd. PerkinElmer got the lowest limit of detection for N gene at 11.61 copies/mL, and the value was 34.66 copies/mL for ORF1ab gene. All of 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.