新型冠状病毒B.1.1.7变异株特异性数字PCR定量方法研究

The Establishment of Digital PCR Quantitative Method for Strain B.1.1.7 of SARS-CoV-2

由于世界卫生组织(WHO)将B.1.1.7(Alpha)列为“关切的变异株”,因此对其建立特异性的高灵敏数字PCR方法尤为重要。该方法需首先根据该变异株刺突蛋白的H69/V70缺失、P681H和A570D 3个特异突变位点设计引物探针,确定3个方法的线性范围和定量限;随后再用其他4个“关切的变异株”验证该方法的特异性,并用不同数字PCR平台和不同方法进行定量准确性的比较;最终,结果表明建立的数字PCR方法准确可靠,可用于B.1.1.7变异株的定量检测。

Due to B.1.1.7(Alpha)being listed as a“variant of concern”by the World Health Organization(WHO),it is particularly important to establish a specific and highly sensitive digital PCR method for it.First,this method required designing primer probes based on the H69/V70 deletion,P681H,and A570D specific mutation sites of the mutant spike protein,and determining the linear range and limit of quantitation for the three methods.Subsequently,the specificity of the method was verified using four other“variant of concern”,and quantitative accuracy was compared using different digital PCR platforms and methods.Finally,the results indicated that the established digital PCR method was accurate and reliable,and can be used for quantitative detection of B.1.1.7 variant strains.