中国寄生虫学与寄生虫病杂志 ›› 2002, Vol. 20 ›› Issue (6): 1-324.

• 论著 •    下一篇

我国多斑按蚊复合体成员种的分子鉴别研究

马雅军;瞿逢伊;董学书;周红宁
  

  1. 第二军医大学病原生物学教研室;第二军医大学病原生物学教研室;云南省寄生虫病防治所;云南省寄生虫病防治所 上海 200433;上海 200433;思茅 665000;思茅 665000
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2002-12-30 发布日期:2002-12-30

Molecular Identification of Anopheles maculatus Complex from China

MA Ya-jun;QU Feng-yi;DONG Xue-shu;ZHOU Hong-ning
  

  1. 1 Department of Etiologic Biology;Second Military Medical University;Shanghai 200433;2 Yunnan Institute of Parasitic Diseases Prevention;Simao 665000
  • Received:1900-01-01 Revised:1900-01-01 Online:2002-12-30 Published:2002-12-30

摘要:   目的建立我国多斑按蚊复合体5成员种的分子鉴别方法。方法测定并分析各成员种的rDNA-ITS2序列,并设计种特异引物,应用PCR法鉴别。结果多斑按蚊复合体5成员种的ITS2序列长度和GC含量分别为伪威氏按蚊328bp、58.54%、多斑按蚊330bp、57.85%、威氏按蚊337bp、59.05%、达罗毗按蚊334bp、58.68%和塞沃按蚊338bp、57.69%,各种内的ITS2序列保守,而种间的差异率范围为9.7%~18.9%。应用5.8S引物和5个种特异引物可以分别扩增出119、186、231、327和406bp等5条清晰不同的种特异条带,以鉴别各蚊种。结论基于ITS2序列差异建立的我国多斑按蚊复合体5成员种的PCR鉴别简便易行、可靠。

关键词: 多斑按蚊复合体, rDNA-ITS2, 分子鉴别

Abstract:  Objective To establish the molecular identification of five members in Anopheles maculatus complex from China. Methods Different rDNA-ITS2 regions of An. maculatus complex were sequenced and analyzed. The species specific primers were designed, and PCR assay was used for the identification. Results The length and GC contents of ITS2 were 328 bp, 58.54% in An. pseudowillmori, 330 bp, 57.85% in An. maculatus, 337 bp, 59.05% in An. willmori, 334 bp, 58.68% in An. dravidicus, and 338 bp, 57.69% in An. sawadwongporni, respectively. The intra-species ITS2 sequences were conservative. The ranges of divergence level among five members were from 9.7% to 18.9% . Five distinct specific fragments were amplified by PCR assay using five species specific primers and 5. 8S primer. The length was 119, 186, 231, 327 and 406 bp respectively. Conclusion The diagnostic PCR assay based on ITS2 divergence to distinguish five members of An. maculatus complex was simple and reliable.

Key words: Anopheles maculatus complex, rDNA-ITS2, molecular identification