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目的确定广东省主要并殖吸虫虫种。方法采用形态学特征观察和分子生物学DNA测序相结合的研究方法,鉴定广东省主要并殖吸虫虫种。采集卫氏并殖吸虫、三平正并殖吸虫和斯氏并殖吸虫自然疫源地山溪中溪蟹并检获三种并殖吸虫囊蚴,分别人工感染家犬或家猫,饲养60~120 d后,剖杀检获成虫分别制成玻片大体标本。结果三种不同并殖吸虫疫源地蟹体检获的囊蚴鉴定为卫氏并殖吸虫、三平正并殖吸虫和斯氏并殖吸虫。三种吸虫成虫样本的COI基因与ITS2基因进行PCR,扩增产物进行测序并分别与Gen Bank中检索的AY618799.1号基因、AF159594.1号基因、AY140693.1号基因、AY618733.1号基因、AF159602.1号基因和AB713404.1号基因序列比对,同源性分别在99%~100%之间。结论广东省主要并殖吸虫为卫氏并殖吸虫、斯氏并殖吸虫和三平正并殖吸虫。三种并殖吸虫的COI基因与ITS2基因序列与Gen Bank中检获的基因序列同源性分别在99%~100%之间,三种并殖吸虫与Gen Bank中检获虫种无明显差异性。
Objective To identify the major Paragonimus species in Guangdong Province. Methods To identify the main Paragonimus species in Guangdong Province by morphological observation and molecular biology DNA sequencing. Collected Paragonimus westermani, Paragonimus spp and Sri Lankan Sri Lanka Paragonimus natural source Creek crab and seized three Paragonimus metacercaria, were infected dog or domestic cats, feeding 60 ~ 120 After d, the seized adults were cut to make slides general specimens. Results The larvae of three species of Paragonimus foetida were identified as Paragonimus westermani, Paragonimus spp. And Paragonimus spp. The COI genes of three species of adult worms were compared with the ITS2 gene and sequenced. The amplified products were sequenced and compared with the AY618799.1, AF159594.1, AY140693.1, AY618733.1 , AF159602.1 gene and AB713404.1 gene sequence alignment, homology were between 99% to 100%. Conclusion The major Paragonimus parasites in Guangdong Province are Paragonimus westermani, Paragonimus spp. The identities of COI gene, ITS2 gene and GenBank in the three Paragonimus strains were between 99% and 100%, respectively. There was no significant difference between the three Paragonimus species and the seized species in GenBank.