新型冠状病毒实时荧光RT-PCR检测的性能验证

Performance verification of a real-time fluorescent RT-PCR for the detection of SARS-CoV-2

目的对实时荧光聚合酶链反应(RT-PCR)方法检测新型冠状病毒(SARS-CoV-2)核酸进行方法学评价.方法依据ISO15189和CNAS-GL039性能评价文件及厂家声明,对本法精密度、准确度、检测下限、特异性和抗干扰能力进行验证.结果RT-PCR试剂检测SARS-CoV-2核酸结果的精密度均小于允许变异度,准确性和重复性的符合率均为100%,检测下限为1.0×103 copies/mL,内源性抗干扰能力CV<5%,与感染部位相同或感染症状相似的其他病原体等无交叉反应.结论RT-PCR检测方法灵敏度高,重复性好,抗干扰力强,检测下限和特异性均符合要求,可用于临床检测.

Objective To evaluate the performance of a real-time fluorescent polymerase chain reaction(RT-PCR)method for the detection of SARS-CoV-2 nucleic acid.Methods To verify the precision,accuracy,lower detection limit,specificity and anti-interference ability of this method,according to ISO 15189 and CNAS-GL039 evaluation documents and manufacturers declaration.Results Precision of RT-PCR reagent of SARS-CoV-2 nucleic acid detection was less than the allowable variability,the accuracy and repeatability coincidence rates were 100%,the lower detection limit was 1.0×10^3 copies/mL,the endogenous anti-interference ability CV was<5%,there was no cross reaction with other pathogens of the same infection site or similar infection symptoms.Conclusions The RT-PCR detection method has high sensitivity,good repeatability,strong anti-interference ability,the lower detection limit and specificity meet the requirements.Therefore,it can be used for clinical detection.