Abstract
The ongoing coronavirus disease 2019 (COVID-19) pandemic has become a public health emergency. Although many reverse-transcription PCR (RT-PCR) assays have been developed, their performance, especially sensitivity assessment, has been insufficiently tested. In this study, a preliminary comparison of the analytical sensitivity of nine RT-qPCR kits from different manufacturers was first conducted using a certified reference material derived from the genomic RNA of SARS-CoV-2 as the template. Subsequently, three of the nine kits, comprising two highly sensitive kits (DAAN, Huirui) and one less sensitive kit (Geneodx), were selected for further sensitivity and specificity validation. The results revealed variations in the performance between kits of the two groups. For the two highly sensitive kits, the limits of detection at 95 % probability (LOD95%) were 5.6 copies of the N gene and 3.5 copies of the ORF 1ab per reaction (DAAN), and 6.4 (N) and 4.6 (ORF 1ab) copies per reaction (Huirui). These LOD95% values were approximately 3 to 4-fold better than those of the Geneodx Kit. However, none of these three Kits showed cross-reactivity against 6 other types of human coronaviruses or respiratory viruses. Because most of these commercial kits are approved as in vitro diagnostics (testing specimens without direct human contact), it would be beneficial for their manufacturers to improve the diagnostic capability of these kits and thus reduce the clinical risks associated with false-negative results.
Keywords: Certified reference material; Limit of detection; RT-qPCR; SARS-CoV-2; Sensitivity.
【저자키워드】 SARS-CoV-2, sensitivity, RT-qPCR, limit of detection, Certified reference material, 【초록키워드】 COVID-19, coronavirus disease, Coronavirus disease 2019, pandemic, Variation, diagnostic, in vitro, RT-PCR, diagnostics, cross-reactivity, respiratory viruses, Probability, sensitivity, RT-qPCR, PCR, Sensitivity and specificity, qPCR, variations, public health emergency, N gene, false-negative results, false-negative, human coronaviruses, analytical sensitivity, genomic RNA, Contact, Limits of detection, reference material, Template, two groups, reaction, specimen, limits, clinical risk, IMPROVE, human coronavirus, selected, tested, approved, conducted, nine, ORF, less, reduce, 【제목키워드】 SARS-CoV-2 detection, validation, nine,