Purpose : This study aims to establish a competitive allele-specific PCR based on penta-primer amplification refractory mutation system (PARMS) technology to detect the key mutation sites of variant of concern (VOC) of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus for rapid typing. Methods : Competitive allele-specific primers and universal primers were designed for the key gene mutation sites N501Y, E484K, L452R, and K417N of SARS-CoV-2 VOCs, respectively.Using the principle of allele-specific polymerase chain reaction and fluorescence energy resonance transfer, different VOCs can be differentiated. Results : Using reverse transcribed cDNA of different VOCs as specimens, through double-blind detection, different VOC types can be effectively identified, with an accuracy rate of 100%. Through the identification and detection of different VOCs, effective differentiation can be achieved. Conclusions : The system has high specificity and sensitivity, with a detection limit of 1.28 copies/reaction.PARMS technology is fast, efficient, and low-cost. It is used for the identification and detection of the popular SARS-CoV-2 VOCs, which is helpful for the rapid and accurate prevention and control of COVID-19 epidemic.
【저자키워드】 COVID-19, SARS-CoV-2, delta variant, variant of concern, rapid typing, 【초록키워드】 coronavirus, Mutation, VoC, variant, virus, amplification, sensitivity, specificity, PCR, Accuracy, L452R, N501Y, K417N, VOCs, E484K, specimens, Detection limit, COVID-19 epidemic, double-blind, cDNA, acute respiratory syndrome, transfer, primer, effective, universal primer, polymerase chain, Result, detect, reverse transcribed, 【제목키워드】 technology, Rapid, strain, typing,