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目的对广西部分地区褐家鼠感染的路氏锥虫进行分子生物学鉴定,分析不同地理种群间的遗传分化。方法从广西靖西、百色、南宁、天峨4地感染路氏锥虫的褐家鼠脾脏或血凝块中提取DNA,设计ITS1保守引物进行PCR扩增,将PCR产物直接或克隆测序。用DNAstar7.1进行同源性比对,用Mega4.0进行差异分析与聚类、构建系统进化树。结果广西各地血涂片阳性鼠标本均扩增出特异性DNA条带,4地样品测序结果与GenBank上登录的路氏锥虫(T.lewisi)序列的同源性均高达96%以上。区内各地理种群之间遗传距离均在0.021以下,与其他3个近缘种T.otospermophili、T.grosi、T.kuseli间遗传距离为0.15～0.206。结论广西4地鼠感染的锥虫为路氏锥虫。ITS1可以作为路氏锥虫的种属分类和分子诊断的标记。
Objective To identify the brown spotted Trypanosoma cruzi in parts of Guangxi by molecular biology and to analyze the genetic differentiation among different geographic populations. Methods DNA was extracted from spleen or blood clots infected with Trypanosoma cruzi from 4 sites in Jingxi, Baise, Nanning, and Tian’e of Guangxi. ITS1 conservative primers were designed for PCR amplification, and the PCR products were directly or cloned and sequenced. DNAstar7.1 homology comparison, Mega4.0 differential analysis and clustering, the construction of phylogenetic tree. Results The specific DNA bands were amplified from the positive smear-positive mice in Guangxi. The results of sequencing showed that all the 4 isolates had the homology of more than 96% with the T. lewisi sequences registered in GenBank. The genetic distances among all geographic populations within the region are below 0.021, and the genetic distances among T.otospermophili, T. grosi and T.kuseli from the other three relatives range from 0.15 to 0.206. Conclusion Trypanosoma Trypanosomiasis in Trypanosoma cartenigensis in Guangxi 4 hamster. ITS1 can serve as a taxonomic classification and molecular marker of Trypanosoma cruzi.