环介导等温扩增技术检测单纯疱疹病毒Ⅰ型的应用

Detection of herpes simplex virus type 2 DNA with a loop-mediated isothermal amplification assay

目的建立一种用于检测单纯疱疹病毒-Ⅰ型(HSV-1)的环介导等温扩增技术(LAMP)检测方法。方法用病毒DNA提取试剂盒提取HSV-1基因组DNA。设计4条扩增HSV-1糖蛋白gG基因的LAMP引物,对HSV-DNA进行LAMP,扩增产物进行琼脂糖电泳和酶切鉴定(试验设HSV-2、HPV-6、HPV-11、沙眼衣原体对照和阴性对照);并对3例临床标本进行LAMP检测。结果 HSV-1 LAMP产物电泳后呈LAMP特征性条带,对照组无扩增条带;扩增产物AvaⅠ酶切片段大小分别为182、140和62 bp,与理论值相符。用HSV-1 LAMP引物扩增3例口唇疱疹标本,2例标本出现LAMP条带,1例标本未扩增出条带。结论建立的LAMP方法简便、高效、快速,可用于HSV-1感染检测。

Objective To detect herpes simplex virus type-1（HSV-1） DNA with a loop-mediated isothermal amplification（LAMP） assay.Methods HSV-1 DNA was extracted with a viral DNA extraction kit.Primers that recognized six regions on the glycoprotein G（gG） gene of HSV-1 were needed and used in LAMP assay.To evaluate the specificity of the assay,sterilized water was used as a negative control.LAMP results were judged by electrophoretic analysis and restriction digestion.Testing featured HSV-2,HPV-6,HPV-11,and Chlamydia Trachomatis controls and a negative control.Three clinical samples were subjected to the LAMP assay.Results LAMP products in the HSV-1 tube were confirmed with gel electrophoresis and a characteristic DNA ladder was observed.In contrast,no bands were observed for the negative control in this study.The specificity of LAMP products was confirmed with restriction enzymes.Of the three samples,two were positive and one was negative.Conclusion A LAMP assay has been established for detection of HSV-1.