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  

Effect of two nucleic acid extraction methods on Zika virus detection by using Real-time RT-PCR

作　　者：刘建礼[1] 刘翌[1] 王飞[1] 田茵[1] 孙福军[1] 乌日嘎[2] 张绍福[1] 祁晓丽[1] 李岩[1] 周燕楠 

机构地区：[1]北京国际旅行卫生保健中心,北京100088 [2]首都机场出入境检验检疫局

出　　处：《中国国境卫生检疫杂志》2016年第3期169-171,179,共4页Chinese Journal of Frontier Health and Quarantine

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.

关 键 词：寨卡病毒 核酸提取 荧光RT-PCR 

分 类 号：R373.3[医药卫生—病原生物学] R466.12[医药卫生—基础医学]