目的  对新型冠状病毒肺炎(COVID-19)康复者恢复期血浆中新型冠状病毒（2019 novel coronavirus，2019-nCoV）核酸检测方法进行系统性验证。方法  根据血浆样本实际情况，并结合试剂盒要求，建立COVID-19康复者恢复期血浆中2019-nCoV核酸的实时荧光定量PCR检测方法，并从专属性、检测限（limit of detection，LOD）、重复性及中间精密度对该方法进行系统性的验证，对20份COVID-19康复者恢复期血浆样本进行性能确认。结果  专属性：阳性样本及阴性样本均能有效被检出；检测限：1×LOD~2×LOD企业参考品19次检测开放阅读框1a/b和核衣壳蛋白基因基因均为阳性；重复性和中间精密度：两种阳性对照基因以及内部质控在不同荧光通道的循环阈值的相对标准偏差均符合要求。性能确认：康复者恢复期血浆2019-nCoV核酸检测结果均为阴性。结论   该检测方法专属性强、稳定可靠、重复性好，适用于COVID-19康复者恢复期血浆2019-nCoV核酸检测。

Objective  To systematically verify the detection method of 2019 novel coronavirus (2019-nCoV)nucleic acid detection in convalescent plasma (CP) samples of COVID-19 patients. Methods  According to actual situation of plasma samples and requirements of the kit, a realtime fluorescent quantitative PCR detection method for 2019-nCoV nucleic acid was established. The method was systematically verified from specificity, detection limit, repeatability and intermediate precision.The method performance was validated by measurements of 20 CP samples. Results  The specificity results showed that both positive and negative samples were detected effectively. The detection limit results revealed that the open reading frame 1a/b and nucleocapsid genes were positive in all of 19 times tests of 1×LOD-2×LOD concentration enterprise reference materials. Repeatability and intermediate precision results showed that the relative standard deviations of cycle threshold values for both positive control genes and internal quality control in different fluorescence channels all met the requirements. The performance validation results displayed that 20 CP samples were all negative for 2019-nCoV nucleic acid. Conclusion  The method is specific, stable, reliable and repeatable, and can be applied for the detection of 2019-nCoV nucleic acid in COVID-19CP samples.