多重核酸即时检测系统在新型冠状病毒和甲/乙型流感病毒联合检测中的应用评价

Application evaluation of multiplexed nucleic acid-POCT method for the detection of SARS-CoV-2,influenza A and B

目的 评价一种基于微流控和实时荧光定量聚合酶链反应(RT-qPCR)结合的技术快速联合检测新型冠状病毒(SARS-CoV-2)、甲型流感病毒(FluA)和乙型流感病毒(FluB)核酸的试剂盒的性能及临床应用。方法 评价新型冠状病毒及甲/乙型流感病毒核酸联合检测试剂盒的灵敏度、特异性、精密度、包容性及抗干扰等性能,并使用临床样本评估本试剂盒在临床应用中的准确性。结果 此试剂盒3种病毒最低检测限均为200 copies/mL;与其他冠状病毒、金黄色葡萄球菌、肺炎链球菌等病原体及人源基因等均没有交叉反应,具有良好的特异性;批内及批间精密度表明不同样本的Ct值变异系数(CV)均≤5%;包容性测试结果显示新型冠状病毒、甲型流感病毒和乙型流感病毒不同亚型的样本均可满足最低检测限及重复性要求;口咽拭子中可能存在的干扰物质,如黏蛋白、苯福林、羟甲唑啉、倍氯米松及地塞米松等对检测结果均无显著影响;临床样本应用一致性评估显示,和传统RT-qPCR方法相比,本试剂盒新型冠状病毒总符合率99.23%,甲型流感病毒阴阳性符合率99.71%,乙型流感病毒阴阳性符合率99.71%。结论 新型冠状病毒、甲型流感病毒和乙型流感病毒核酸联合检测试剂盒性能良好,适用于多种场景即时快速检测需求。

Objective To evaluate the performance and clinical application of point-of-care testing(POCT)kits based on microfluidics and real-time quantitative polymerase chain reaction for rapid joint detection of SARS-CoV-2,influenza A and B.Methods The sensitivity,specificity,precision,inclusiveness and anti-interference performance of the joint detection kit for SARS-CoV-2,influenza A and B were evaluated,and the accuracy of this kit in clinical application was evaluate using the clinical samples.Results The minimum detection limit of three viruses in this system was 200 copies/mL.It had no cross-reaction with other kinds of coronavirus,staphylococcus aureus,streptococcus pneumoniae,other pathogens and human genes.The intra assay and inter assay precision showed that the coefficient of variation of Ct values of different samples were≤5%.The results of inclusive test showed that the minimum detection limit and repeatability of different sub-types of SARS-CoV-2,influenza A and influenza B were consistent.The possible interfering substances in the oropharynx swab,such as mucin,phenylephrine,hydroxymethylzoline,beclomethasone,dexamethasone,etc.,had no significant impact on the testing results.The evaluation on the consistence of clinical samples application showed that,compared with the traditional RT-qPCR method,the total coincidence rate of this kit for SARS-CoV-2 was 99.23%,the coincidence rate for influenza A was 99.71%,and the coincidence rate for influenza B was 99.71%.Conclusions This joint detection kit for nucleic acids of SARS-CoV-2,influenza A virus and Influenza B virus has good performance and is suitable for real-time and rapid detection in multiple scenarios.