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目的：建立一种鉴别大劣按蚊复合体Ａ和Ｄ种的ＰＣＲ检测技术。方法：根据大劣按蚊Ａ和Ｄ种核糖体ＤＮＡ第二内转录间隔区的序列差异，设计种特异引物，通过ＰＣＲ扩增出种特异长度（Ａ种为３７４ｂｐ，Ｄ种为６６３ｂｐ）的片段区分蚊种。结果：该法敏感性达１／１６００单蚊抽提ＤＮＡ或１／５单条蚊腿水匀浆ＤＮＡ。应用该法检测大劣按蚊ＡＦＲＩＭＳ实验室品系１０例，ＨＮ实验室品系２０例，以及采自海南省白沙、和平、罗葵、毛阳等地野外样本１４８例，均显示Ａ种特异带；云南省勐腊地区野外样本３０例，全部为Ｄ种特异带。结论：提供一种简便可靠的蚊种鉴别ＰＣＲ法，确认我国海南和云南的大劣按蚊分别为Ａ种和Ｄ种，为深入研究我国大劣按蚊复合体不同成员种的地理分布、生态习性和传疟作用创造了条件。
OBJECTIVE: To establish a PCR assay to identify Anopheles dirus complex A and D species. Methods: Species-specific primers were designed according to the sequence differences of the second internal transcribed spacer region of Anopheles dirus A and D ribosomal DNA. The species-specific primers (374 bp in type A and 663 bp in type D) were amplified by PCR Distinguish between mosquito species. Results: The method was sensitive to 1/1600 mosquitoes extracted DNA or 1/5 mosquito and leg homogenate DNA. The method was applied to detect 10 AFRIMS laboratory strains, 20 HN laboratory strains and 148 wild samples from Baisha, Heping, Luo Kui and Mao Yang in Hainan Province, all showing A specific bands. Thirty cases of wild samples in Mengla area of Yunnan Province were all D specific bands. Conclusion: To provide a simple and reliable method to identify mosquito species, confirm that Anopheles dirus in Hainan and Yunnan are A species and D species respectively. In order to further study the geographical distribution and ecological features of different species of Anopheles dirus in China, Habits and the role of malaria created the conditions.