哈氏并殖吸虫ITS2基因和CO1基因序列分析

目的 进一步确认浙江宁海哈氏并殖吸虫（Paragonimus harinasutai）分类地位及其遗传变异。 方法 从浙江华溪蟹中分离获得哈氏并殖吸虫囊蚴，单个囊蚴经PCR扩增，进行核糖体DNA第二间区（ITS2）基因和线粒体细胞色素C氧化酶亚单位（CO1）基因序列分析；ITS2-PCR扩增产物经BsaHI、StuI酶切。 结果 ITS2-PCR扩增产物经BsaHI和StuI酶切（PCR-RFLP），琼脂糖凝胶电泳分离图谱基本一致。浙江宁海哈氏并殖吸虫ITS2基因有366碱基对，与泰国种群（AF159609）核酸同源性为95.6％，CO1基因有390碱基对，与泰国种群（AF159600）核酸同源性为89.5％。 结论 本研究从分子生物学角度进一步确认浙江宁海哈氏并殖吸虫分类地位，但与泰国种群（Nakorn-nayok省）相比较，存在较大变异。

Sequence Analysis of ITS2 and CO1 Genes of Paragonimus harinasutai

Objective To identify Paragonimus harinasutai from Ninghai,Zhejiang Province,China.Methods Meta-cercariae were collected from the crabs Sinopotamon chekiangenes in Xixi village of Ninghai County for ITS2 sequence analysis,CO1 sequence analysis and endonuclease BsaHI and StuI analysis by PCR-RFLP.Results The fingerprintings of PCR-RFLP were virtually same to the isolate from Thailand(Nakorn-nayok).The ITS2 sequence with 366 bp and CO1 sequence with 390 bp of the metacercariae collected from Ninghai revealed a nucleotide identity 95.6% and 89.5% respectively to the Thai isolate.Conclusion The study confirmed that Paragonimus harinasutai is present in Ninghai,China,with certain variation on molecular biology in comparison to the Thai isolate.