多重核酸检测试剂盒RespiFinder■ 2SMART在检测12种呼吸道感染病毒中的临床应用性能评价

Evaluation on clinical performance of a multiplex nucleic acid detection kit RespiFinder■2SMART in detection of 12 viruses in respiratory tract infections

目的评价多重核酸检测试剂盒RespiFinder■2SMART在临床呼吸道感染病原体检测中的性能。方法选取2012年5月至2015年5月青海省发热呼吸道症候群病例哨点医院收集的471份鼻咽拭子作为研究材料,采用单管多重核酸检测试剂盒RespiFinder■2SMART及实验室自建方法对12种病毒进行平行检测,分析两种方法的检测结果一致性。对于不一致的样本采用巢式PCR结合测序或单重荧光定量PCR方法进行分析。以实验室检测结果为参照,分析试剂盒的阳性预测值、阴性预测值、灵敏度、特异度及约登指数等指标,综合评估试剂盒的性能。结果两种方法对12种呼吸道病毒的检出符合率均超过95.00%。对结果不一致的样本(10.20%,48/471)复核,证实了试剂盒检出的HCoV-229E、HAdV和HRV/HEV部分样品为真阳性(38.00%,23/48)。以实验室检测结果为参考标准,试剂盒对大部分病原检出的阳性预测值和阴性预测值均可达到100.00%,但检出HCoV-229E和HAdV的阳性预测值偏低(50.00%和87.93%,95%CI为2.56~97.44和77.13~94.03);试剂盒对大部分病原体检出的灵敏度和特异度均可达到100.00%,但检出PIV-1、PIV-2和RSV-A的灵敏度偏低(58.33%,57.14%,64.28%,95%CI为31.95~80.67,99.17~100.00,38.76~83.00);试剂盒检测大部分病原的约登指数均超过0.98,但检测PIV-1、PIV-2和RSV-A的约登指数分别为0.58、0.57和0.64。结论RespiFinder■2SMART试剂盒用于临床样本中HCoV、HMPV、HAdV、HRV、PIV-3及RSV-B的检测性能好,但对于PIV-1,PIV-2和RSV-A的检测灵敏度还有待提高。

Objective To evaluate the performance of a commercial multiplex nucleic acid detection kit, i.e. RespiFinder■ 2SMART kit in the pathogen detection of respiratory infections in clinical practice.Methods From May 2012 to May 2015, 471 nasopharyngeal swabs specimens collected from patients with acute febrile respiratory syndrome in Qinghai province were used as the research materials. The single tube multiplex nucleic acid detection kit, i.e. RespiFinder■ 2SMART kit and the self-made methods established in the laboratory were used in parallel to detect 12 viruses. The consistency rates of both methods were analyzed. The samples with discordant results were analyzed by nested PCR and sequencing or by fluorescent quantitative PCR with single set of primers. The performance of the kit was evaluated by accessing positive predictive value, negative predictive value, sensitivity, specificity and Youden's index of the kit with the results by the self-made methods as reference values.Results The consistency rates for all targeted viruses between the two methods were higher than 95.00%. For the samples with discordant results (10.20%, 48/471), the positive results of HCoV-229E, HAdV and HRV/HEV by the kit (38.00%, 23/48) were confirmed to be correct. Using the results by the self-made methods as reference values, the positive predictive values and negative predictive values of the kit were 100.00% for most targeted viruses while the positive predictive values were relatively low for HCoV-229E (50.00%, 95%CI: 2.56-97.44) and HAdV (87.93%, 95%CI: 77.13-94.03). Both sensitivities and specificities of the kit were 100% for most targeted viruses while the sensitivities were low for PIV-1 (58.33%, 95%CI: 31.95-80.67), PIV-2 (57.14%, 95%CI: 99.17-100.00) and RSV-A (64.28%, 95%CI: 38.76-83.00). The Youden’s indexes of the kit for most targeted viruses were higher than 0.98 while the indexes of PIV-1, PIV-2, and RSV-A were 0.58, 0.57 and 0.64, respectively.Conclusions The RespiFinder■ 2SMART kit demonstrated good performance in detection of HCoV, HMPV, HAdV, HRV, PIV-3 and RSV-B, but the sensitivity should to be improved for detection of PIV-1, PIV-2 and RSV-A.