分子分类结合形态学观察鉴定重庆地区片形吸虫种类

CHARACTERISATION OF FASCIOLA IN CATTLE AND BUFFALO FROM CHONG QING USING ITS MEDIATED PCR AND MORPHOUS METHODS

目的明确重庆地区片形吸虫的种类,并为重庆地区片形吸虫的分类研究提供科学的参考依据。方法在对重庆地区黄牛、水牛肝脏上寄生的片形吸虫形态结构进行观察后,根据片形吸虫第一内转录间隔区(ITS-1)和第二内转录间隔区(ITS-2)基因设计特异性引物,运用多聚酶链式反应(PCR)技术,以法国肝片吸虫gDNA为对照,对所采样品gDNA用大片吸虫和肝片吸虫的ITS-1和ITS-2特异性引物进行扩增。结果形态学鉴定均为“不规则”片形吸虫,电泳结果显示均分别扩增出特异性ITS-1和ITS-2条带。结论综合形态学和PCR鉴定结果,初步认为重庆地区存在着大片吸虫和肝片吸虫的“中间型”。

Objective To characterize the Fasciola sp and to provide the proof for the classification of Fasciola sp in ChongQing. Methods The Fasciola. sp in cattle and buffalo was handled and observed, then the polymerase chain reacation（PCR） was applied according to the primers which were designed based on the ITS-1 and ITS- 2 sequences of Fasciola hepatica and Fasciola gigantica respectively, and the gDNA of Fasciola hepatica from France as the positive control. Results The morphological analysis displayed the two kinds of Fasciola was neither the standard type of Fasciola hepatica nor Fasciola. gigantica, and the two special lanes of ITS-1 and ITS-2 were also showed in Fasciola hepatica and Fasciola gigantica respectively. Conclusion the Fasciola in Cattle and buffalo in Chang Qing is more probably the ＂intermediate＂ Fasciola and it has provided the proof for the classification of Fasciola sp.