通用引物检测登革热病毒NS1基因及其酶切分型  

Detection and Typing of Dengue Viruses by RT-PCR with Universal Primers Coupled with Restriction Enzyme Analysis

作　　者：舒莉萍[1] 左丽[1] 李永念[1] 郝牧[1] 陈阿英[1] 

机构地区：[1]贵阳医学院免疫学教研室,贵州贵阳550004

出　　处：《贵阳医学院学报》2004年第4期283-286,共4页Journal of Guiyang Medical College

基　　金：国家自然科学基金资助项目 (39960 0 0 1 )

摘　　要：目的 :利用通用引物经逆转录 -聚合酶链反应 (RT PCR)检测登革病毒RNA ,并经限制性内切酶酶切鉴定病毒型别。方法 :用C6 / 36细胞培养登革病毒 ,碘化钠法提取病毒RNA ,利用通用引物进行RT PCR ,对PCR产物用MavⅠ限制性内切酶酶切鉴定。结果 :应用通用引物RT PCR方法可检测到含量为 0 2 1 2ng/L的病毒RNA ,扩增的产物及酶切片段与预期的靶序列长度和限制酶谱分析相一致。结论 :用登革病毒NS1区通用引物通过RT PCR并经酶切分型 。Objective: To diagnose and classify dengue virus. Methods: A pair of universal primers was designed according to NS1 gene of dengue virus was used in the reverse transcriptase polymerase chain reaction(RT PCR) to diagnose dengue virus. Dengue virus was cultured with C6/36 cells. The total RNA was extracted. The viral NS1 gene fragment was amplified with RT PCR, and the products were digested with MavⅠfor typing. Results: DEN 1 Hawaii strain RNA was amplified from infected tissue culture fluid at concentration as low as 0.212 ρg/ml. Conclusions: This pair of primers is charactered by universality and this method reflects good quality of sensitivity and specificity.

关 键 词：登革热病毒 RNA 逆转录聚合酶链反应 DNA限制酶类 

分 类 号：R373.33[医药卫生—病原生物学]