聚合酶链式反应(PCR)扩增基因鉴定蚊胃血血源

Identification of Mosquito Gastric Blood Sources Through Gene Amplification by Polymerase Chain Reaction( PCR)

目的探寻鉴定蚊胃血来源的实验室快速检测方法。方法对抗人血红蛋白胶体金试纸条(简称试纸条)检测有人血成分的按蚊干血痕标本,依据常见蚊吸血对象(人、牛、猪)的DNA序列的差异,设计特异DNA基因引物,运用聚合酶链式反应(PCR)方法对按蚊胃内干血痕标本进行DNA基因扩增,1.5%琼脂糖电泳检测扩增产物,并对扩增产物进行序列测定后查询其同源性匹配。结果试纸条共检测87只按蚊,血源来自牛和猪的共83只,检出仅含人血痕4份。该4份人血痕蚊标本经PCR扩增检测和基因序列匹配查询,与人基因序列同源性为100%,而无牛和猪基因。结论PCR法鉴定蚊胃血血源快速、灵敏、特异。

Objective To explore a rapid detection method for identification of mosquito gastric blood sources in lab. Methods Immuno -colloidal gold test strips was adopted to detect the dried blood stains of Anopheles containing human blood and peculiar DNA gene primer was designed according to the differences of DNA se- quences of common blood - sucking objectives, such as human, cattle and swine, thereby PCR was used to amplify the DNA sequences, which was detected with 1.5% agarose eleetrophoresis; finally, the amplifica- tion products were determined and conducted homology matching. Results Totally 87 Anopheles were tested, the gastric blood of 83 were belonged to cattle and swine and four belonged to human. After PCR amplifica- tion test and gene sequence matching, the sequence homology of four blood stains was 100% with human＇ s and none with cattle or swine＇ s. Conclusion The PCR method is of rapidity, sensitivity and specificity in i- dentifying the mosquito gastric blood sources.