新型冠状病毒核酸快速检测系统质量评估

Quality assessment of rapid nucleic acid detection system for the novel coronavirus

目的本研究旨在评估快速检测系统对新型冠状病毒(SARS-CoV-2)核酸项目检测能力和检测质量,为快速检测系统的应用和结果的分析提供一定的参考依据。方法选用SARS-CoV-2假病毒和RNA合成全片段两种类型的能力验证质控品作为阳性样本,人冠状病毒229E灭活样本作为阴性样本。使用SARSCoV-2 RNA合成全片段能力验证质控品和阴性样本验证快速检测系统的性能,包括符合率、精密度和最低检测限,比较快速检测系统和常规PCR设备的精密度和检出能力,以及探索快速检测系统对低于最低检测限浓度样本检出能力;选用这两种类型的阳性样本评估一步法和常规磁珠法提取SARS-CoV-2 RNA后经快检设备扩增对结果的影响。结果快速检测系统的符合率为100.00%;快速检测系统和常规PCR设备检测两个阳性浓度水平样本的精密度CV值均小于5.00%,对浓度500 copy/mL样本的检出率为100.00%;两种类型的阳性样本经磁珠法与一步法提取核酸后,在快检设备上扩增检测,两者结果比较,经磁珠法提取的样本检测结果的ORF1ab和N基因的Ct值均较低,差异有统计学意义(P<0.05);快速检测系统对浓度为400 copy/mL样本检出率为100.00%,对浓度为200 copy/mL样本N和ORF1ab基因检出率分别为100.00%和91.67%,对浓度为100 copy/mL样本N和ORF1ab基因检出率分别为91.67%和58.33%。结论该快速检测系统对新型冠状病毒核酸检测性能良好,但常规磁珠法提取SARS-CoV-2 RNA比该快检设备厂家推荐的一步法提取检测的灵敏度更高。

Objective To evaluate the detection ability and detection quality of the rapid detection system for novel coronavirus(SARS-CoV-2)nucleic acid items,and provide a certain reference for the clinical applica-tion and the results analysis of the rapid detection system.Methods Two types of proficiency testing sam-ples,such as SARS-CoV-2 pseudovirus and RNA synthesis complete fragments,were selected as positive sam-ples,and human coronavirus 229E inactivated samples were used as negative samples.Using SARS-CoV-2 RNA-positive samples and negative samples to verify the performance of the rapid detection system,including coincidence rate,precision and limit of detection(LOD),and to compare the precision and detection capability of the rapid detection system and conventional PCR equipment.Besides,we wana explore the ability of rapid detection systems to test samples with concentrations below the LOD.These two types of positive samples were chosen to assess the effect of one-step and conventional magnetic bead extraction on the results of rapid detection equipment amplification of SARS-CoV-2 RNA.Results The coincidence rate of the rapid detection system is 100.00%.The precision CV values of the two positive concentration level samples tested by the rap-id detection system and conventional PCR equipment were both less than 5.00%,and the detection rate of the sample with a concentration of 500 copies/mL was 100.00%.The two types of positive samples were ampli-fied and detected on the rapid detection equipment after nucleic acid extraction by magnetic bead method and one-step method.Compared with these results,the Ct values of ORF1ab and N genes of the former were low-er,and the difference was statistically significant(P<0.05).The detection rate of the rapid detection system was 100.00%for samples with a concentration of 400 copies/mL,100.00%and 91.67%for the N and ORF1ab genes for samples with a concentration of 200 copies/mL.The detection rates of N and ORF1ab genes for samples with a concentration of 100 copies/mL were 91.67%and 58.33%,respectively.Conclusion The rapid detection system manifests well at the detection of new coronavirus nucleic acid,but the extraction of SARS-CoV-2 RNA by the conventional magnetic bead method is more sensitive than the one-step extraction which was recommended by the manufacturer of the rapid detection equipment.