Analytical performance evaluation of five RT‐PCR kits for severe acute respiratory syndrome coronavirus 2. Issue 1 (27th October 2020)

• Record Type: Journal Article
• Authors: Wang, Mengmeng; Chen, Dayang; Wu, Wei; Tang, Huamei; Kan, Lijuan; Zong, Zengyan; Dou, Xiaowen; Ji, Xiang; Xiong, Dan; Zhang, Xiuming
• Is Part Of: Journal of clinical laboratory analysis. Volume 35:Issue 1(2021)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2020-10-27
• Subjects: analytical performance -- detection ability -- limit of detection -- repeatability and imprecision -- RT‐PCR -- SARS‐CoV‐2; Diagnosis, Laboratory -- Periodicals; Medical laboratory technology -- Periodicals; 616
• DOI: 10.1002/jcla.23643
• Languages: English
• ISSNs: 0887-8013

Abstract

Abstract: Background: We aimed to evaluate the analytical performance of five commercial RT‐PCR kits (Genekey, Daan, BioGerm, Liferiver, and Yaneng) commonly used in China, since such comparison data are lacking. Methods: A total of 20 COVID‐19 confirmed patients and 30 negative nasopharyngeal swab specimens were analyzed by five kits. The detection ability of five RT‐PCR kits was evaluated with 5 concentration gradients diluted by a single positive sample. The limit of detection was evaluated by N gene fragment solid standard. Two positive clinical specimens were used to evaluate the repeatability and imprecision. Finally, we used six human coronaviruses plasmid and four respiratory pathogens plasmid to check for cross‐reactivity. Results: The positive detection rate was 100% for Genekey, Daan, and BioGerm, and 90% for Liferiver and Yaneng in 20 clinical SARS‐CoV‐2 infection. The coincidence rate of five kits in 10 negative samples was 100%. The detection rate of target genes for Daan, BioGerm, Liferiver, and Yaneng was 100% from Level 1 to Level 3. In Level 4, only Daan detection rate was 100%. In Level 5, five kits presented poor positive rate. The limit of detection declared by each manufacturer was verified. The repeatability for target genes was less than 5% and so did the total imprecision. There is no cross‐reactivity of five kits with six human coronaviruses and four respiratory pathogens for ORF1ab and N gene. Conclusions: Five RT‐PCR kits assessed in this study showed acceptable analytical performance characteristics and are useful tools for the routine diagnosis of SARS‐CoV‐2. Abstract : In the work, we presented the analytical performance evaluations of five RT‐PCR kits using nasopharyngeal swabs samples from patients with confirmed SARS‐CoV‐2 infection, and negative nasopharyngeal swabs samples (Figure 1). A total of 20 COVID‐19 confirmed patients and 30 negative nasopharyngeal swab specimens were analyzed by five kits. The detection ability of five RT‐PCR kits was evaluated with 5 concentration gradients diluted by a single positive sample. The limit of detection was evaluated by N gene fragment solid standard. Two positive clinical specimens were used to evaluate the repeatability and imprecision. Finally, we used six human coronaviruses plasmid and four respiratory pathogens plasmid to check for cross‐reactivity