新冠病毒核酸检测试剂盒的性能分析与评价

Performance Analysis and Evaluation of SARS-CoV-2 Nucleic Acid Test Kits

目的:对4种新型冠状病毒核酸检测试剂盒的性能包括敏感性与特异性、最低检出限、精密度和分析特异性进行评价,为实验室选择新冠病毒核酸检测试剂盒提供参考,促进新冠肺炎病毒核酸检测技术的创新改进。方法:以20例新冠病毒确诊阳性和10例确诊阴性的咽拭子作为研究样本,经提取得到RNA后根据4种新冠病毒核酸检测试剂盒说明书对其进行检测,分析扩增结果,评估各试剂盒的差异。结果:检测结果显示4种试剂盒对30例研究样本的阴阳性判定情况不尽相同,其中a试剂盒的敏感性和特异性最高,均达到100%;3例阳性样本稀释到100倍时,c试剂盒只能检出其中的1例,而其他三种试剂盒能检出其中的2例,表明c试剂盒的检测敏感性低于其他三种;4种试剂盒在精密度实验中检测样本得到的Ct值的CV值都较接近,且都<10%;加入了8种常见的高浓度干扰物质的临界阳性样本经4种试剂盒检测,结果依然为弱阳性,说明无干扰;4种试剂盒检测5例含非目标病毒的临床样本,检测结果为阴性,即未发生交叉反应。结论:新冠病毒核酸检测试剂盒对低浓度核酸样本的检测能力有所差异,各试剂盒不同的检出限也造成了总体上的敏感性和特异性的差异,实验室在选择新冠病毒核酸检测试剂盒时可以对其进行性能验证,以保证选到最合适的试剂盒。

Objective:To evaluate the performance of four novel Coronavirus nucleic acid test kits,including sensitivity and specificity,minimum detection limit,precision and analysis specificity,to provide reference for laboratory selection of SARS-CoV-2 nucleic acid test kits and promote the innovation and improvement of SARS-CoV-2 nucleic acid detection technology.Methods:Twenty SARS-CoV-2 positive and ten novel Coronavirus negative pharyngeal swabs were used as research samples.RNA was extracted and detected according to the instructions of four SARS-CoV-2 nucleic acid test kits.The amplification results were analyzed and the differences of each kit were evaluated.Results:The test results showed that the negative and positive determination of the four kits on the 30 samples were different,among which the sensitivity and specificity of a kit were the highest,reaching 100%;When 3 positive samples were diluted to 100 times,the c kit could only detect 1 of them,while the other three kits could detect 2 of them,indicating that the sensitivity of the c kit was lower than that of the other three;The CV values of Ct values obtained by the four kits in the precision test were all close,and all were less than 10%;The critical positive samples with 8 kinds of common high concentration interfering substances were tested by 4 kinds of kits,and the results were still weak positive,indicating no interference.5 clinical samples containing non-target viruses were tested with four kits,and the test results were negative,that is,no cross-reaction occurred.Conclusion:SARS-CoV-2 nucleic acid test kits differ in their ability to detect low-concentration nucleic acid samples,and the LOD of each kit also results in differences in overall sensitivity and specificity.When selecting novel Coronavirus nucleic acid test kits,laboratories can verify their performance to ensure that the most appropriate kit is selected.