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328 bp、58.54％、多斑按蚊330 bp、57.85％、威氏按蚊337 bp、59.05％、达罗毗按蚊334 bp、58.68％和塞沃按蚊338 bp、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406 bp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。
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.
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
2002年第6期321-324,共4页
Chinese Journal of Parasitology and Parasitic Diseases
基金
国家自然科学基金(No.39670663
39900127)
