核酸快速检测系统在新型冠状病毒检测中的应用评价

目的:探讨核酸快速检测系统在新型冠状病毒(SARS-CoV-2)检测中的性能与应用评价。方法:收集北京市疾病预防控制中心和中日友好医院检验科2020年2至7月的临床样本,评价核酸快速检测系统对SARS-CoV-2检测的敏感度、特异度、抗干扰能力、精密度以及临床样本检测符合率。分析敏感度的评价通过检测浓度梯度稀释的SARS-CoV-2样本,每个样本重复检测20次;分析特异度的评价是使用该系统检测与SARS-CoV-2感染部位相同或症状相似的病原体或假病毒及人源DNA和RNA;抗干扰能力研究是通过检测具有潜在干扰物质的样本进行评价;精密度研究通过评价试剂批内及批间的检测差异;临床样本检测符合率研究使用该系统检测230例咽拭子样本和95例痰液样本,并与传统实时荧光定量PCR(RT-qPCR)结果及临床诊断结果比较。结果:核酸快速检测系统对SARS-CoV-2核酸检测的分析敏感度为400拷贝/ml;核酸快速检测系统对感染部位相同或感染症状相似的病原体和人源核酸的检测结果均为阴性,分析特异度表明该系统与相关病原体及人源核酸无交叉反应;精密度表明批内/批间试验的高、中、低浓度样本的Ct值变异系数为1.90%~3.92%,均小于5%;加入潜在干扰物质后样本检测结果仍为阳性,说明样本中可能存在的干扰物质对检测结果无影响;与RT-qPCR结果及临床诊断结果符合率分别为98.46%和97.85%。结论:基于分子并行反应的核酸快速检测系统可作为一种快速检测SARS-CoV-2的选择方法。

Evaluation of fast nucleic acid detection system in severe acute respiratory syndrome coronavirus 2

Objective To evaluate the performance and application of a fast nucleic acid detection system for testing severe acute respiratory syndrome virus 2(SARS-COV-2).Methods Clinical samples were collected from February to July 2020 from Beijing Center for Diseases Prevention and Control and the Laboratory Department of China-Japan Friendship Hospital,to evaluate the sensitivity,specificity,anti-interference ability,precision and clinical sample coincidence rate of fast nucleic acid detection system for SARS-CoV-2.The analytical sensitivity was determined by a dilution series of 20 replications for each concentration.Analytical specificity study was performed by testing organisms whose infection produces symptoms similar to those observed at the onset of corona virus disease 2019(COVID-19),and of the normal or pathogenic microflora that may be present in specimens collected.Potential interference substances were evaluated with different concentration in the interference study.Precision study was conducted by estimating intra-and inter-batch variability.Clinical evaluation was performed by testing 230 oropharyngeal swab specimens and 95 sputum specimens in fast nucleic acid detection system,comparing with conventional real-time fluorescent quantitative PCR(RT-qPCR)and clinical diagnostic results.Results The analytical sensitivity of SARS-CoV-2 using fast nucleic acid detection system was 400 copies/ml.The result is negative for testing with the organisms that may likely in the circulating area or causing similar symptoms with SARS-CoV-2 and human nucleic acid,indicating that no cross reactivity with organisms.The results of precision test showed that the Coefficient of variation of Ct value of high,medium and low concentration samples was 1.90%-3.92%,and all of them were less than 5%in intra-and inter-batch testing.The results of the samples were still positive after adding the potential interfering substances,indicating that the possible interfering substances in the samples had no effect on the results.98.46%and 97.85%diagnosis results of fast nucleic acid detection system were consistent with RT-qPCR and clinical diagnostic results,respectively.Conclusion The fast nucleic acid detection system based on molecular parallel reaction can be used as a selection method for SARS-CoV-2 testing.