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

Q4 Pharmacology, Toxicology and Pharmaceutics

国际药学研究杂志 Pub Date : 2020-06-30 DOI:10.13220/J.CNKI.JIPR.2020.06.004

C. Yang, W. Liu, L. Qu, Y. Ke, Y. Chen, R. Jia, X. Liu, Y. Wang, L. Han, L. Jia, C. J. Wang

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引用次数: 0

Abstract

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 ex- traction 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

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临床标本SARS-CoV-2核酸检测试剂盒及核酸提取方法的比较评价

目的：比较分析7种新型冠状病毒核酸检测试剂盒和5种核酸提取方法方法；选取33份新冠病毒阳性临床样本进行核酸提取方法比较。采用5种不同的核酸提取方法提取样本，然后进行RT-PCR扩增实验，比较阳性率和Ct值。结果：A品牌核酸提取试剂盒阳性率最高，漏检率最低；核酸提取方法的比较表明，手动柱提取法阳性率最高，其次是磁珠提取法，一步提取法漏检率最高。结论：严重急性呼吸系统综合征冠状病毒2型检测试剂盒的检测能力参差不齐，因此在选择试剂盒之前需要进行评估工作。手动柱提取法显示出最佳的提取效率，但耗时较长。由于可以与自动核酸提取仪相结合，磁珠提取法具有较高的提取效率，该方法可能适用于大批量样品的提取。尽管一步提取法易于操作，但该方法的漏检率较高，因此不建议临床使用该方法

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来源期刊

国际药学研究杂志 Pharmacology, Toxicology and Pharmaceutics-Pharmaceutical Science

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发文量

10973

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