PCR-RFLP和特异PCR技术用于鲁道夫对盲囊线虫鉴定的研究

Determination of specific status of Contracaecum rudolphii from China by PCR-RFLP and Specific PCR assays

本研究运用新建立的Contracaecum rudolphii姊妹种的PCR-RFLP和特异PCR鉴定方法对来自我国青海湖的鲁道夫对盲囊线虫进行分子鉴定。经PCR-RFLP分析,来自我国青海湖的鲁道夫对盲囊线虫的两个样品的带型与C.rudolphii B的酶切带型一致。经特异PCR扩增,C.rudolphii B的特异引物能从我国鲁道夫对盲囊线虫中扩增出目的片段,与对照C.rudolphii B的代表样品扩增片段大小相同。该两种方法试验结果都说明来自我国青海湖的对盲囊线虫为C.rudolphii B,与PCR-SSCP方法鉴定结果一致。本次研究结果及所建立的分子分类学方法为我国异尖线虫的进一步研究奠定了基础。

The present study was designed to determine the specific status of specimens representing Contracaecum rudolphii, collected from Qinghai Lake in China, by PCR-linked restriction fragment length polymorphism （PCR-RFLP） and specific PCR assays. The result of PCR-RFLP analysis revealed that the restriction profiles of C. rudolphii specimens from China were identical to that of C. rudolphii B. Specific amplicons were amplified from the Chinese C. rudolphii specimens using the C. rudolphii B-specific primers, identical in lengh to that of C. rudolphii B. These findings indicated that C. rudolphii specimens from China represented C. rudolphii B. The findings of the presented study and the two molecular assays taken have implications for studying the systematics and molecular genetics of anisakid nematodes from China.