4种全自动核酸提取系统对新型冠状病毒假病毒提取效果的比较

耿帜; 徐远东; 包一熙; 陈凤花

doi:10.13201/j.issn.1004-2806.2021.10.001

4种全自动核酸提取系统对新型冠状病毒假病毒提取效果的比较

- 华中科技大学同济医学院附属协和医院检验科(武汉, 430022)

详细信息

通讯作者: 陈凤花,E-mail:chfh100@126.com

中图分类号: R446.1

收稿日期: 2021-03-15

Comparative evaluation of four automated nucleic acid extraction platforms using 2019-nCoV pseudovirus

- Department of Clinical Laboratory, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China

More Information

Corresponding author: CHEN Fenghua, E-mail: chfh100@126.com

Received Date: 15 March 2021

摘要

摘要: 目的：应用4种全自动核酸提取仪及其配套试剂提取新型冠状病毒（2019-nCoV）假病毒核酸，并用同1种商品化RT-PCR扩增试剂盒进行检测，探讨检测结果的差异以评估其提取效果，有助于选择合适的核酸提取平台。方法：将2019-nCoV假病毒连续梯度稀释成不同浓度，用4种不同厂家的核酸提取系统（包括全自动核酸提取仪及其配套提取试剂） A、B、C和D提取核酸，再用同1种2019-nCoV核酸检测试剂盒进行检测，比较其检出率和循环阈值（Ct）的差异。结果：在2019-nCoV假病毒原浓度、1：5、1：10和1：100稀释度时，A、B和D提取的核酸中均能100%检测到ORF1ab和N基因，进一步分析发现这3种平台提取的核酸中检测所得的ORF1ab （F=0.061，P=0.941）和N基因（F=0.038，P=0.963） Ct值差异均无统计学意义；但C在2019-nCoV假病毒1：100稀释时所提取的核酸中ORF1ab和N基因的阳性检出率分别为95%和80%。结论：平台C与其他3种平台A、B、D的核酸提取效果间存在差异。核酸提取平台的不同可能影响2019-nCoV核酸检测的灵敏度，并且可能对病毒载量低的样本产生假阴性结果。
- 核酸 /
- 提取 /
- 新型冠状病毒假病毒
Abstract: Objective: To compare and evaluate four different automated magnetic bead-based nucleic acid extraction instruments and paired kits for purifying nucleic acids from 2019-nCoV pseudovirus.Methods: Nucleic acids were extracted from serial dilutions of 2019-nCoV pseudovirus using four nucleic acid extraction platforms A, B, C and D, and detected by the commercial 2019-nCoV nucleic acid detection kit.Results: Both ORF1 ab and N genes were detected in the nucleic acids extracted from 2019-nCoV pseudovirus and serial dilutions including 1:5, 1:10 and 1:100 by A, B and D, and there was no significant difference between cycle threshold(Ct) values of ORF1 ab(F=0.061, P=0.941) and N genes(F=0.038, P=0.963) detected by the same amplification kit in the nucleic acids extracted by A, B and D; but the positive detection rates of ORF1 ab and N genes in the nucleic acids extracted from 1:100 dilution of 2019-nCoV pseudovirus using C were 95% and 80%, respectively.Conclusion: The platform C was different from the other three platforms A, B and D in the nucleic acid extraction. The nucleic acid extraction platform may influence the sensitivity of detecting 2019-nCoV and have the potential to generate false-negative results especially in the samples with low viral load.
- nucleic acid /
- extraction /
- 2019-nCoV pseudovirus

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参考文献(5)

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