用核糖体DNA鉴别PCR区分中华按蚊和嗜人按蚊  

Differentiation of Anopheles sinensis and Anopheles anthropophagus using a ribosomal DNA PCR assay

作　　者：马雅军[1,2] 瞿逢伊[1,2] 徐建农[1,2] 郑哲民 

机构地区：[1]第二军医大学基础医学部寄生虫学教研室 [2]陕西师范大学生命科学学院

出　　处：《第二军医大学学报》1998年第3期237-239,共3页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金

摘　　要：目的：建立鉴别中华按蚊和嗜人按蚊的ＰＣＲ检测技术，并对现场标本进行检测。方法：根据中华按蚊和嗜人按蚊的ｒＤＮＡＩＴＳ２序列差异，设计种特异引物，用ＰＣＲ技术扩增种特异ＤＮＡ片段（中华按蚊４２５ｂｐ，嗜人按蚊２５３ｂｐ），根据扩增片段的长度对两种按蚊进行鉴别。结果：应用该法检测中华按蚊和嗜人按蚊的３个实验室品系共７０例，经卵鉴定的嗜人按蚊野外标本２０例，均显示单一的种特异扩增带。凉山按蚊、贵阳按蚊和帕氏按蚊对照共９例全部无扩增。检测采自我国１０省１２个不同地点，根据成蚊形态初步鉴定为中华按蚊的标本共４４０例，具中华按蚊种特异扩增带的标本占６６．１％，具嗜人按蚊种特异扩增带的标本占１２．７％，其余２１．１％的标本无扩增带，表明检测样本中混有其他蚊种，仅根据成蚊形态难以鉴别。结论：本研究提供了一种简便、准确和灵敏的中华按蚊和嗜人按蚊的分子鉴别方法。Objective: To develop a diagnostic PCR assay for distinguishing Anopheles sinensis and An. anthropophagus, and to test its usefulness for detecting field specimens. Methods: The primers of the PCR assay were designed based on the sequence difference in ribosomal DNA internal transcribed spacer 2 of An. sinensis and An. anthropophagus. The diagnostic lengths of species specific fragments were 425 bp in An. sinensis and 253 bp in An.anthropophagus. Results: For a total of 70 specimens of An. sinensis and An. anthropophagus laboratory lines and 20 field specimens of An. anthropophagus identified by egg characters, the PCR assay gave expected speciesspecific amplified fragments. At the presence of DNA from An.

关 键 词：中华按蚊 嗜人按蚊 DNA 核糖体 PCR 

分 类 号：R384.1[医药卫生—医学寄生虫学]