新型冠状病毒核酸血液筛查方法的验证及应用

Validation and application of a blood screening method for SARS-CoV-2 nucleic acid

目的对适用于新型冠状病毒(severe acute respiratory syndrome coronavirus 2,SARS-CoV-2)核酸血液筛查的检测系统进行性能评价和临床应用,为SARS-CoV-2核酸检测在新冠康复者血浆筛查中的应用提供依据和技术手段。方法采用实时荧光定量PCR法检测SARS-CoV-2核酸,对95%检出限(LOD)、精密度、抗干扰等指标进行验证,同时应用该检测系统对新冠疫情期间采集的献血者标本进行SARS-CoV-2核酸筛查,评价其适用性。结果该检测方法对N基因和ORF 1ab基因检出限分别为3.98 copies/mL、9.38 copies/mL。检测高浓度、低浓度标本的变异系数均<5%。500 mg/dL的游离血红蛋白以及3 g/dL的甘油三酯对检测结果影响小。同时检测系统可有效防止携带污染,避免假阳性结果。献血者SARS-CoV-2核酸血液筛查过程中,共计检测39306份血液标本均呈阴性。结论该核酸血液筛查检测方法可满足血液中SARS-CoV-2核酸筛查的需求,保障新冠肺炎康复者血浆的应用安全性。

Objective To evaluate the performance and clinical application of a high-throughput nucleic acid blood screening detection system for SARS-CoV-2,so as to provide basis and technical means for its application in detection of plasma from recovered COVID-19 patients.Methods The SARS-CoV-2 nucleic acid was detected by real-time fluorescence quantitative PCR,and the sensitivity,precision,anti-interference and other parameters were evaluated.Blood donor samples collected during COVID-19 epidemic period were screened using the detection system to evaluate its applicability.Results The detection limits of gene N and ORF 1ab were 3.98 copies/mL and 9.38 copies/mL,respectively.The CV of high and low concentration samples were both less than 5%.Hemoglobin at 500 mg/dL and triglyceride at 3 g/dL had little effect on the results.The detection system can effectively prevent carryover,thus avoiding false positive results.The SARS-CoV-2 nucleic acid blood screening was carried out in a total of 39306 blood samples,and all samples were negative.Conclusion The established method can meet the needs of SARS-CoV-2 nucleic acid screening therefore ensure the safe application of plasma from recovered COVID-19 patients.