新型冠状病毒样本核酸检测及核酸提取方法效果评价

Comparative evaluation of nucleic acid detection kits and nucleic acid extraction methods for SARS-CoV-2 using clinical samples

目的对7种新型冠状病毒核酸检测试剂盒以及5种核酸提取方法进行比较分析。方法选取44例新型冠状病毒阳性临床样本,核酸提取后利用7种新型冠状病毒核酸检测试剂盒进行RT-PCR扩增实验,比较不同试剂盒的阳性率及Ct值;选取33例新型冠状病毒阳性临床样本,利用5种核酸提取方法对样本进行提取,后对提取的核酸进行RT-PCR扩增实验,比较不同试剂盒的阳性率及Ct值。结果核酸提取试剂盒中品牌A的阳性率最高,漏检率最低;核酸提取方法比较显示,手工过柱法提取的核酸阳性率最高,其次为磁珠法,一步法检测的漏检率最高。结论目前新型冠状病毒检测试剂盒的检测能力参差不齐,在选用之前需做好评价工作。核酸提取方法中,手工过柱法提取效果好但耗时较长;磁珠法由于能配套核酸自动提取仪使用,故提取效率高,适合大批量样本的提取工作;一步法虽步骤简便,但漏检率高,不建议临床使用。

Objective To compare and analyze 7 new coronavirus nucleic acid detection kits and 5 nucleic acid extraction methods.Methods After extracting nucleic acids from 44 positive coronavirus clinical samples,7 SARS-CoV-2 nucleic acid detection kits were used for RT-PCR amplification experiments to compare the positive rate and Ct value;33 new coronavirus positive clinical samples were selected to compare the acid extraction methods.Five different nucleic acid extraction methods were used to extract the samples,and then RT-PCR amplification experiments were performed to compare the positive rate and Ct value.Results The brand A nucleic acid extraction kit had the highest positive rate and the lowest rate of missed detection;comparison of nucleic acid extraction methods showed that the manual column extraction method had the highest positive rate,followed by the magnetic bead extraction method,and the one-step extraction method had the highest missed detection rate.Conclusion The detection capabilities of the SARS-CoV-2 detection kits are uneven,so evaluation work needs to be done before the selection of the kit.The manual column extraction method showed best extraction efficiency but took a long time.Because of the possible combination with the automatic nucleic acid extraction instrument,the magnetic bead extraction method had a high extraction efficacy,which might be suitable for use in the extraction of large batches of samples.Although the one-step extraction method was easily operable,the method had a high missed detection rate,so this method was not recommended for clinical use.