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浙江大学学报(医学版)  2020, Vol. 49 Issue (2): 185-190    DOI: 10.3785/j.issn.1008-9292.2020.03.09
原著     
严重急性呼吸综合征冠状病毒2(SARS-CoV-2)核酸检测试剂盒临床诊断效能评估
沈利华1(),黄菲2(),陈祥1,熊钻3,杨晓玉2,李慧2,程丰2,郭健1,*(),龚国富2,*()
1. 湖北省鄂州市妇幼保健院检验科, 湖北 鄂州 436000
2. 湖北省鄂州市中心医院检验科, 湖北 鄂州 436000
3. 湖北省鄂州市鄂钢医院检验科, 湖北 鄂州 436000
Diagnostic efficacy of three test kits for SARS-CoV-2 nucleic acid detection
SHEN Lihua1(),HUANG Fei2(),CHEN Xiang1,XIONG Zuan3,YANG Xiaoyu2,LI Hui2,CHENG Feng2,GUO Jian1,*(),GONG Guofu2,*()
1. Department of Clinical Laboratory, Ezhou Maternal and Child Health Hospital, Ezhou 436000, Hubei Province, China
2. Department of Clinical Laboratory, Ezhou Central Hospital, Ezhou 436000, Hubei Province, China
3. Department of Clinical Laboratory, Egang Hospital, Ezhou 436000, Hubei Province, China
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摘要:

目的: 了解三种严重急性呼吸综合征冠状病毒2(SARS-CoV-2)核酸检测试剂盒的检测效果及临床诊断效能。方法: 采集40例临床诊断为2019冠状病毒病(COVID-19)和16例非COVID-19住院患者的咽拭子样本。采用湖南圣湘生物科技有限公司(以下简称“圣湘公司”)、硕世生物科技股份有限公司(以下简称“硕世公司”)、深圳华大基因股份有限公司(以下简称“华大基因”)SARS-CoV-2核酸RT-PCR检测试剂盒检测上述样本并分析其敏感度、特异度、阳性预测值、阴性预测值和Kappa值。比较一步裂解法和磁珠法、两种不同核酸提取仪(圣湘公司Natch CS S12C全自动核酸提取系统和西安天隆科技有限公司NP968-C核酸提取仪)磁珠法提取咽拭子样本中病毒核酸的效果及其RT-PCR检测结果的阳性符合率、阴性符合率、总符合率及Kappa值。结果: 圣湘公司试剂盒检测SARS-CoV-2核酸的敏感度、特异度、阳性预测值、阴性预测值和Kappa值分别为95.00%、87.50%、95.00%、87.50%和0.825,硕世公司试剂盒分别为90.00%、87.50%、94.74%、77.78%和0.747,华大基因试剂盒分别为82.50%、81.25%、91.67%、65.00%和0.593。一步裂解法和磁珠法提取的病毒核酸检测结果阳性符合率、阴性符合率、总符合率和Kappa值分别为95.24%、100.00%、96.43%、0.909,但一步裂解法仅耗时25 min,而磁珠法需180 min。两种不同核酸提取仪磁珠法提取的RNA检测结果阳性符合率、阴性符合率、总符合率和Kappa值分别为85.00%、100.00%、89.29%和0.764。结论: 圣湘公司试剂盒检测SARS-CoV-2核酸效果略优于其他两家公司试剂盒。一步裂解法提取咽拭子样本中总RNA具有操作简单、省时和检测结果符合率较高的优点。

关键词: 2019冠状病毒病严重急性呼吸综合征冠状病毒2新型冠状病毒肺炎诊断技术聚合酶链反应诊断效能    
Abstract:

Objective: To compare the diagnostic efficacy among three RT-PCR test kits for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleic acid detection. Methods: The throat swab samples from 40 hospitalized patients clinically diagnosed as coronavirus disease 2019 (COVID-19) and 16 hospitalized non-COVID-19 patients were recruited. The SARS-CoV-2 nucleic acid was detected in throat swab samples with RT-PCR test kits from Sansure Biotech ("Sansure" for short), Jiangsu Bioperfectus Technologies ("Bioperfectus" for short) and BGI Genomics ("BGI" for short). The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and Kappa value were analyzed. The viral nucleic acid was extracted from the throat swab samples by one-step cleavage and magnetic bead methods, and the efficacy of two extraction methods was also compared. The results of magnetic bead method for nucleic acid extraction by two different extractors (Sansure Natch CS S12C Fully Automated Nucleic Acid Extraction System vs. Tianlong NP968-C Nucleic Acid Extractor) were also compared. Results: The sensitivity, specificity, PPV, NPV and kappa value were 95.00%, 87.50%, 95.00%, 87.50%and 0.825 for Sansure kit; 90.00%, 87.50%, 94.74%, 77.78%and 0.747 for the Bioperfectus kit, and 82.50%, 81.25%, 91.67%, 65.00%and 0.593 for the BGI kit, respectively. The positive, negative and total coincident rates and kappa value of viral nucleic acid detection results using the samples extracted by one-step cleavage and magnetic bead methods were 95.24%, 100.00%, 96.43%and 0.909, respectively, but the one-step cleavage method took only 25 min, while the magnetic bead method required 180 min. The positive, negative and total coincident rates and kappa value of viral nucleic acid detection results using the samples extracted by the two different nucleic acid extractors were 85.00%, 100.00%, 89.29% and 0.764, respectively. Conclusion: The detection efficacy for SARS-CoV-2 nucleic acid by the Sansure kit is relatively higher and the one-step cleavage method has advantages of convenient operation and less time consuming.

Key words: Coronavirus disease 2019    Severe acute respiratory syndrome coronavirus 2    Novel coronavirus pneumonia    Diagnostic techniques    Polymerase chain reaction    Diagnostic efficacy
收稿日期: 2020-03-05 出版日期: 2020-03-17
:  R446  
通讯作者: 郭健,龚国富     E-mail: 4458629@qq.com;416672475@qq.com;252895179@qq.com;402389961@qq.com
作者简介: 沈利华(1984-), 男, 学士, 主管技师, 主要从事病原微生物检测研究; E-mail:4458629@qq.com; https://orcid.org/0000-0002-4436-2770|黄菲(1980-), 女, 学士, 主管技师, 主要从事免疫学检验研究; E-mail:416672475@qq.com; https://orcid.org/0000-0002-5053-7442
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引用本文:

沈利华,黄菲,陈祥,熊钻,杨晓玉,李慧,程丰,郭健,龚国富. 严重急性呼吸综合征冠状病毒2(SARS-CoV-2)核酸检测试剂盒临床诊断效能评估[J]. 浙江大学学报(医学版), 2020, 49(2): 185-190.

SHEN Lihua,HUANG Fei,CHEN Xiang,XIONG Zuan,YANG Xiaoyu,LI Hui,CHENG Feng,GUO Jian,GONG Guofu. Diagnostic efficacy of three test kits for SARS-CoV-2 nucleic acid detection. J Zhejiang Univ (Med Sci), 2020, 49(2): 185-190.

链接本文:

http://www.zjujournals.com/med/CN/10.3785/j.issn.1008-9292.2020.03.09        http://www.zjujournals.com/med/CN/Y2020/V49/I2/185

试剂盒 靶基因 核酸提取 加样量
(μL)
扩增温度和循环数结果判断最低检测限
(拷贝数/mL)
内标
阳性 阴性
  SARS-CoV-2:严重急性呼吸综合征冠状病毒2;Ct值:每个反应管内的荧光信号到达设定阈值时所经历的循环数.
圣湘公司 ORF1ab/N 磁珠法
一步裂解法
10
20
60 ℃ 45个循环 S型扩增曲线
Ct值≤40
Ct值>40 200
硕世公司 ORF1ab/N 磁珠法 5 55 ℃ 45个循环 S型扩增曲线
Ct值≤35
Ct值>38 1000
华大基因 ORF1ab 磁珠法 10 60 ℃ 40个循环 S型扩增曲线
Ct值≤38
不呈S型曲线
且无Ct值
100
表 1  三种SARS-CoV-2核酸检测试剂盒的基本情况和技术指标
试剂盒COVID-19样本(n=40)非COVID-19样本(n=16)
阳 性 阴 性 阳 性 阴 性
  SARS-CoV-2:严重急性呼吸综合征冠状病毒2;COVID-19:2019冠状病毒病.
圣湘公司 38 2 2 14
硕世公司 36 4 2 14
华大基因 33 7 3 13
表 2  三种SARS-CoV-2核酸检测试剂盒的临床诊断效能
总RNA
提取方法
COVID-19样本(n=40)非COVID-19样本(n=16)
阳 性 阴 性 阳 性 阴 性
  COVID-19:2019冠状病毒病.
一步裂解法 40 0 2 14
磁珠法 38 2 2 14
表 3  一步裂解法与磁珠法检测效果比较
核酸
提取仪
COVID-19样本(n=40)非COVID-19样本(n=16)
阳 性 阴 性 阳 性 阴 性
  COVID-19:2019冠状病毒病.
圣湘公司 38 2 2 14
天隆公司 33 7 1 15
表 4  不同核酸提取仪对检测效果的影响
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