新标准下17种新型冠状病毒核酸检测试剂性能比对研究

Comparative study on the performance of seventeen 2019-ncov acid detection reagents under the new standard

目的参照新型冠状病毒肺炎诊疗方案(试行第九版)中解除集中隔离和出院核酸检测标准比较17种新型冠状病毒核酸检测试剂的检测性能,为实验室核酸检测选择试剂提供参考。方法选取2021-08/2022-03新型冠状病毒肺炎疑似病例呼吸道样本50份,经数字聚合酶链式反应(dPCR)检测确定样本阴阳性并对病毒核酸载量定量,再分别对17种检测试剂(编号为A~Q)进行评价。以Ct值35为限值判定检出数和检出率,利用各试剂的检测值做试剂间两两比对分析,对各试剂检检测值和dPCR检测值比对分析Ct值与核酸量的关系,以P<0.05为差异有统计学意义。结果17种核酸检测试剂中15种试剂检测正常,阴性样本检出率在0%~35%之间,阳性样本检出率在39%~96%之间,各试剂间检出率和检测值均有一定差异且与检测靶标数、试剂检测最低限以及检测模板加入量没有相关性;核酸量在57360 copies/ml以上各试剂均能检出,且检出能力随核酸量减少出现差异。结论若检测结果不按照试剂盒说明书中Cut-off值而是按照Ct值35为限值判定,则15种试剂对较高病毒载量的样本均能有效检出,但各试剂的检测性能有一定差异,因此在核酸检测工作中应配备多种品牌检测试剂以相互验证提高检测准确率。

Objective To compare the detection capacity of 172019-nCoV nucleic acid testing reagents according to the nucleic acid test standards for lifting centralized isolation and discharging from hospital refer to COVID-19 diagnosis and treatment Protocol(Trial Edition 9),and to provide reference for the selection of nucleic acid test reagents in laboratory.Methods Fifty respiratory tract samples from COVID-19 suspected patients from 2021-08/2922-03 were selected.The samples were divided into negative and positive by digital polymerase chain reaction(dPCR)to evaluated 17detection reagents(numbered A-Q)respectively.Ct value 35 was used as the limit to determine the number of detections and detection rate,and the detection value of each reagent was used for pairwise comparison between the reagents.The relationship between Ct value and nucleic acid content was analyzed by comparing the detection value of each reagent and dPCR detection value.P<0.05 was considered as statistically significant difference.Results 15 of the 17 nucleic acid detection reagents were work.The detection rate of negative samples was 0%~35%,and the detection rate of positive samples was between 39%~96%.There were some differences in the detection rate and detection value of each reagent,and there was no correlation with the number of detection targets,the minimum detection limit of reagent and the amount of detection template.All the reagents can be detected with nucleic acid content above 57360copies/mL,and the detection ability varies with the decrease of nucleic acid content.Conclusions If the deteccion results are determined according to Ct value 35 instead of Cut-off value in the kit instrctions,all the 15reagents can effectively detect the samples with high viral load detection performance of each reagent is different to some extent.Therefore,multiple brands of detection reagents should be equipped in nucleic acid detection to verify each other and improve the detection accuracy.