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目的 分析和比较不同地区中华按蚊和嗜人按蚊的ITS2区段的基因特征。方法 采用特异性ITS2引物对嗜人按蚊和中华按蚊江苏实验株以及从湖北省和越南现场捕获的嗜人按蚊和中华按蚊进行PCR扩增、克隆并对ITS2区段序列进行分析。结果 嗜人按蚊实验株的ITS2区段序列有452bp,与嗜人按蚊现场株的ITS2区段序列相同,中华按蚊实验株的ITS2区段序列有472bp,与中华按蚊现场株的ITS2区段序列也相同;但嗜人按蚊和中华按蚊的ITS2区段基因序列存在明显差异并存在不同的限制性内切酶位点。结论 可依据中华按蚊和嗜人按蚊的ITS2基因序列内限制性内切酶切位点不同的基因特征,采用PCR-RFLP技术建立中华按蚊和嗜人按蚊基因鉴别技术。
Objective To analyze and compare the genetic characteristics of ITS2 in Anopheles sinensis and Anopheles anthropophagus in different areas. Methods Specific ITS2 primers were used to amplify the ITS2 segments of Anopheles anthropophagus and Anopheles sinensis Jiangsu experimental strains, and Anopheles anthropophagus and Anopheles sinensis captured from Hubei province and Vietnam. Results The sequence of ITS2 of Anopheles anthropophagus was 452bp, which was the same as the ITS2 sequence of Anopheles anthropophagus. The sequence of ITS2 of Anopheles sinensis was 472bp, which was close to the ITS2 The sequence of the ITS2 segment of Anopheles anthropophagus and Anopheles sinensis is also significantly different and there are different restriction enzyme sites. Conclusion According to the different gene characteristics of restriction endonuclease sites in the ITS2 gene sequence of Anopheles sinensis and Anopheles anthropophagus, PCR-RFLP technique was used to identify the Anopheles sinensis and Anopheles anthropophagus genes.