Analytical performance evaluation of five RT‐PCR kits for severe acute respiratory syndrome coronavirus 2

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.

Recurrence, Reactivation, or Inflammatory Rebound of SARS-CoV-2 Infection With Acute Vestibular Symptoms: A Case Report and Revision of Literature

“…They include false RT-PCR results or positive due to traces of the RNA genome, intermittent virus shedding, viral reactivation in people with low antibody levels or immunity, reinfection with another SARS-CoV-2 strain, an acute severe systemic inflammatory response known as cytokine release syndrome (CRS), or exposure to a contaminated environmental surface after discharge (Yang et al, 2020 ; Dao et al, 2021 ). Various molecular diagnostic assays have been developed and used worldwide, but the differences in their diagnostic performances remain poorly understood (Matsumura et al, 2020 ; Liotti et al, 2021 ; Wang M. et al, 2021 ). Most of the articles do not report the commercial kit used for RT-PCR, and, where reported, the sensitivity and specificity data for the kit is not often available in the literature ( Table 1 ).…”

Diagnostic performance of a colorimetric RT -LAMP for the identification of SARS-CoV-2: A multicenter prospective clinical evaluation in sub-Saharan Africa

“…Extracted RNA was tested by RT-PCR with the novel coronavirus (2019-nCoV) real-time multiplex RT-PCR kit (Liferiver #RR-0479-02, Revision No. ZJ0010 produced in China) [ 15 , 16 ] and by the SARS-CoV-2 rapid colorimetric LAMP assay kit (New England Biolabs #E2019S produced in US) [17] . The Liferiver RT-PCR targets three gene loci separately in the SARS-CoV-2 genome: Orf1ab, E and N, while the RT-LAMP from New England Biolabs targets E and N simultaneously.…”

Epidemiological feature, viral shedding, and antibody seroconversion among asymptomatic SARS-CoV-2 carriers and symptomatic/presymptomatic COVID-19 patients

“…Quantitative reverse transcription-PCR (qRT-PCR) assay was applied to detect SARS-CoV-2 genomic RNA in nasal and throat swabs, sputum, and feces of patients. Patients in Ningbo were examined using the test kits manufactured by Shanghai BioGerm Medical Technology (Shanghai, China) and Daan Gene Co., Ltd (Guangzhou, China) [13]. Patients in Zhoushan were examined using the test kits manufactured by Shanghai GeneoDx Biotech Company (Shanghai, China) [14].…”