Comparative performance of four nucleic acid amplification tests for SARS-CoV-2 virus

Yujuan Xiong; Zhen-Zhen Li; Qizhen Zhuang; Yan Chao; Fei Li; Yi-Yuan Ge; Yi Wang; Pei-Feng Ke; Xianzhang Huang

amplification tests (NAATs) have been rapidly developed and quickly applied to 23 clinical testing during the pandemic. However, studies evaluating the performance of 24 these NAAT assays are limited. We evaluated the performance of four NAATs, which 25 were marked by the Conformité Européenne and widely used in China during the 26 pandemic. Results showed that the analytical sensitivity of the four assays was 27 significantly lower than that claimed by the NAAT manufacturers. The limit of 28 detection (LOD) of Daan, Sansure, and Hybribio NAATs was 3000 copies/mL, 29 whereas the LOD of Bioperfectus NAATs was 4000 copies/mL. The results of the 30 consistency test using 46 samples showed that Daan, Sansure, and Hybribio NAATs 31 could detect the samples with a specificity of 100% (30/30) and a sensitivity of 100% 32 (16 /16), whereas Bioperfectus NAAT detected the samples with a specificity of 100% 33 (30/30) and a sensitivity 81.25% (13/16). The sensitivity of Bioperfectus NAAT was 34 lower than that of the three other NAATs; this finding was consistent with the result 35 that Bioperfectus NAAT had a higher LOD than the three other kinds of NAATs. The 36 four above mentioned reagents presented high specificity; however, for the detection 37 of the samples with low virus concentration, Bioperfectus reagent had the risk of 38 missing detection. Therefore, the LOD should be considered in the selection of Introduction 46 Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has spread rapidly 47 since its recent identification in patients with severe pneumonia in Wuhan, China(1-3). 48 To date, SARS-CoV-2 has affected more than 372,000 patients worldwide and 49 resulted in more than 16,000 deaths (as of March 24, 2020)(4). The World Health 50 Organization (WHO) has declared the Chinese outbreak of coronavirus disease 2019 51 to be a Public Health Emergency of International Concern and stated 52 that the spread of COVID-19 may be interrupted by early detection, isolation, prompt 53 treatment, and the implementation of a robust system(4, 5). On March 11, at a regular 54 press conference in Geneva, WHO Director-General Tan Desai said that the 55 COVID-19 epidemic can be called a pandemic in terms of its characteristics.