七种国产新型冠状病毒核酸检测试剂盒的一致性和检出能力评价研究

Evaluation of the consistency and detection capability of seven domestic 2019-nCoV nucleic acid detection kits

目的比较7种国产新型冠状病毒(2019-nCoV)核酸检测试剂盒的一致性和检出能力,为临床实验室选择检测方法和新型冠状病毒肺炎的诊断提供参考依据。方法收集深圳市罗湖区人民医院2020年1月29日至2月5日2019-nCoV感染确诊患者核酸检测阳性和疑似患者核酸检测阴性的咽拭子标本各10例,采用7种试剂盒(编号a^g)分别进行核酸检测,评价7种试剂盒临床标本检测结果的一致性;选取1份阳性标本核酸用无RNA酶水梯度稀释得到5个浓度梯度盘(浓度1~浓度5),比较不同品牌试剂盒的阳性检出率及批内重复性。结果6种试剂盒检测20例临床标本的阴性和阳性符合率均为100%,仅1种试剂盒的阳性符合率为8/10,阴性符合率10/10;批内重复性显示7种试剂盒在浓度1水平重复检测病毒载量CV值均<5%;在浓度1~浓度3梯度区间,开放阅读框(ORF)1ab基因的检出能力比较显示b、d、f低于a、c、e、g,试剂盒e和g最高(14/15),a试剂盒对N基因检出能力可达15/15,优于其余5种试剂盒。ORF1ab和N基因检出能力综合分析显示d试剂盒检出能力最低(ORF1ab:40%;N:53%),a、b、c、e、f这5种试剂盒检出能力无明显差异。结论7种试剂盒对病毒载量较高的阳性标本检测的准确度和重复性无明显差异,检测性能良好;但部分试剂盒对弱阳性标本的检出能力欠佳,建议弱阳性标本应至少采取两个厂家的试剂盒复核检验,以保障结果的准确性。

Objective To compare the consistency and detection capability of seven 2019-nCoV nucleic acid detection kits,and provide reference for detection method selection of clinical laboratory and diagnosis of new coronavirus pneumonia.Methods Two batches of pharyngeal swab samples were collected from tenpatients with confirmed infection of 2019-nCoV and 10 suspected patients with negative 2019-nCoV test results during January 29 to February 5,2020 in Shenzhen Luohu People′s Hospital.Seven kinds of kits were labeled as ato g and used for nucleic acid detection respectively to evaluate the consistency of the test results of the clinical samples.A 2019-nCoV positive specimen was selected and diluted to 5-concentration gradient plates(Level-1 to 5)with RNase-free water.The positive detection rate and intra-batch repeatability of different brands of kits were compared.Results The negative and positive coincidence rates of twenty clinical samples tested by six kinds of kits were 100%,and the positive and negative coincidence rate was 8/10 and 10/10 for the other kit,respectively.The results of intra-batch repeatability showed the CVs of viral loads tested by these seven kits were all less than 5%.In the concentration range of Level-1 to 3,the detection capability for open reading frame(ORF)1ab gene of Kit b,d and f was lower than Kit a,c,e and g,and the detection capability of kit e and g was the highest(14/15).The detection capability for N gene of Kit a(15/15)was higher than the other 5 kits.The comprehensive analysis of the detection capability for ORF1ab and N gene showedthat Kit d had the lowest detection capability(ORF1ab:40%,N:53%),and there was no significant difference in the detection capability of Kit a,b,c,e,and f.Conclusions There was no significant difference in the accuracy and repeatability of the seven kits for positive samples with high viral loads,and the detection performance was good;but some kits had poor detection capability for weak positive samples.It is suggested that the weak positive samples should be rechecked by at least two manufacturers′kits to ensure the accuracy of the results.