两种核酸提取方法在寨卡病毒实时荧光RT-PCR检测中的比较
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摘要
目的比较硅胶膜法和磁珠法在寨卡病毒核酸检测中的使用效果,以选择适合的核酸提取方法用于临床样本的寨卡病毒检测。方法对北京首例输入性寨卡病毒感染病例的尿液和唾液样本同时用硅胶膜法和磁珠法提取总RNA,用一步法和两步法实时荧光RT-PCR进行寨卡病毒核酸检测,比较检测效果的差异。结果在一步法中,用硅胶膜法从尿液中提取RNA,寨卡病毒检测阳性;用磁珠法从尿液和唾液中提取的RNA,检测均为阴性。在二步法中,用硅胶膜法从尿液和唾液中提取的RNA,检测均为阳性,而磁珠法提取的样本均未检出。结论硅胶膜法提取核酸比磁珠法更适合于寨卡病毒荧光RT-PCR检测。
Objective To compare the performance of two commonly used nucleic acid extraction methods,silica membranes column and magnetic beads,in the detection of Zika virus detection. Methods Total RNA were extracted from urine and saliva samples,which had been collected from the first imported case of Zika virus infection in Beijing,by silica membranes column and magnetic beads,respectively. One-step or two-step Real-time RT-PCR were conducted to detect Zika virus,and the performance of two extraction methods were compared.Results RNA extracted from urine sample treated by silica membranes was detected positive in one-step Real-time RT-PCR,while samples treated by magnetic beads were all negative. In two-step Real-time RT-PCR,RNA were detected positive from both urine and saliva samples extracted by silica membranes,but undetected in all samples extracted by magnetic beads. Conclusion Nucleic acid extraction by silica membranes was more effective for Zika virus detection by using Real-time RT-PCR.
Objective To compare the performance of two commonly used nucleic acid extraction methods,silica membranes column and magnetic beads,in the detection of Zika virus detection. Methods Total RNA were extracted from urine and saliva samples,which had been collected from the first imported case of Zika virus infection in Beijing,by silica membranes column and magnetic beads,respectively. One-step or two-step Real-time RT-PCR were conducted to detect Zika virus,and the performance of two extraction methods were compared.Results RNA extracted from urine sample treated by silica membranes was detected positive in one-step Real-time RT-PCR,while samples treated by magnetic beads were all negative. In two-step Real-time RT-PCR,RNA were detected positive from both urine and saliva samples extracted by silica membranes,but undetected in all samples extracted by magnetic beads. Conclusion Nucleic acid extraction by silica membranes was more effective for Zika virus detection by using Real-time RT-PCR.
引文
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[11]李成德,黄晓佳,陈雄毅.不同核酸提取方法在HBV DNA荧光定量检测中的比较[J].国际检验医学杂志,2010,31(12):1361-1363.
[12]龚黎明,葛琼,卢亦愚,等.4种核酸提取方法在流行性感冒病毒核酸检测中的比较[J].中国卫生检验杂志,2007,17(1):53-54.
[2]中国疾病预防控制中心.寨卡病毒实验室检测技术方案.[EB/OL].[2016-3-29].http://www.chinacdc.cn/jkzt/crb/qt/ablcxr_8561/zstd_6217/201603/t20160329_128203.html.
[3]北京市卫生和计划生育委员会.北京市报告1例输入性寨卡病毒病病例[EB/OL].[2016-5-15].http://www.bjhb.gov.cn/wsxw/201605/t20160515_137133.html.
[4]Gourinat AC,O'Connor O,Calvez E,et al.Detection of Zika Virus in Urine[J].Emerg Infect Dis.2015,21(1):84-86.
[5]Lanciotti RS,Kosoy OL,Laven JJ,et al.Genetic and Genetic and serologic properties of Zika virus associated with an epidemic,Yap State,Micronesia,2007[J].Emerg Infect Dis.,2008,14(8):1232-1239.
[6]王森,马学军.核酸提取方法的研究进展[J].中华实验与临床病毒学杂志,2014,28(6):503-505.
[7]吴斌,李彩东,段正军,等.磁珠法在HCV RNA定量检测中的临床应用[J].国际检验医学杂志,2014,35(15):2090-2091.
[8]何世成,林源,王昌建,等.五种核酸提取法对猪繁殖与呼吸综合征病毒提取效率的比较[J].中国动物检疫,2014,31(8):70-72.
[9]张险朋,董瑜,温贤诚.三种核酸提取方法检测禽流感病毒的比较[J].动物医学进展,2014,35(11):116-119.
[10]谭国浩,谭翰清,姬宏超.不同核酸提取试剂在肠道病毒核酸检测中的比较[J].中国热带医学,2015,15(12):1543-1545.
[11]李成德,黄晓佳,陈雄毅.不同核酸提取方法在HBV DNA荧光定量检测中的比较[J].国际检验医学杂志,2010,31(12):1361-1363.
[12]龚黎明,葛琼,卢亦愚,等.4种核酸提取方法在流行性感冒病毒核酸检测中的比较[J].中国卫生检验杂志,2007,17(1):53-54.