绒山羊和绵羊源东毕吸虫ITS rDNA的PCR扩增及序列分析

PCR Amplification and Sequence Analysis of the ITS rDNA of Orientobilharzia spp. from Goat and Sheep in Hei Longjinang Province

目的扩增绒山羊和绵羊源东毕吸虫的ITS rDNA序列并进行分析比较。方法运用PCR方法,以保守引物BD1和BD2扩增从黑龙江绒山羊和绵羊体内分离的东毕吸虫(Orientobilharzia spp.)rDNA的内转录间隔区(ITS-1及ITS-2)。PCR产物经测序后进行序列分析。结果黑龙江绒山羊和绵羊东毕吸虫的ITS序列总长均为875bp,其中ITS-1序列长为384bp,5.8S序列长为159bp,ITS-2序列长为332bp。黑龙江绒山羊和绵羊东毕吸虫的ITS序列相似性为99.9%,只有一个碱基差异,存在于ITS-2序列中。结论本研究在国际上首次报道了绒山羊和绵羊东毕吸虫的ITS序列,证实绒山羊和绵羊源东毕吸虫代表同一个种。这些结果为东毕吸虫分子生物学的进一步研究奠定了基础,并为寄生虫分子系统学研究提供了新资料。

Objective The aim of the present study was to obtain the ITS sequences of Orientobilharzia spp. from goat and sheep in Hei Longjiang province. Methods The DNA region spanning the first internal transcribed spacer （ITS-1）, 5.8S and the second internal transcribed spacer （ITS-2） of ribosomal DNA was amplified by PCR using a pair of conserved primers from individual samples from goat and sheep in Hei Longjiang province in China. The amplicons were sequenced directly and compared with the relevant sequences. Results The length of ITS-1, 5.8S and ITS-2 representing the two Orientobilhazia samples was 384 bp, 159 bp, and 332 bp, respectively. There was only a single base difference in the ITS-2 between the sample from goat and sheep. Conclusion Results from the present study demonstrate that Orientobiharzia from sheep and goat represent the same species.