浙江宁海哈氏并殖吸虫的形态学和分子生物学

Paragonimus Harinasutai from Ninghai,Zhejiang:Morphology and Molecular Biology

目的进一步确认浙江宁海哈氏并殖吸虫分类地位。方法1.形态学观察:浙江华溪蟹采自浙江宁海西溪冷风洞。分离囊蚴感染家猫,成虫布氏液固定,洋红染色,制成整体封片标本作形态观察。2.分子生物学研究:单个囊蚴PCR扩增,进行ITS2基因和CO1基因序列分析,并BsaHI、StuI酶切。结果成虫长叶型,宽长之比为1:2.838±0.228。腹吸盘位于虫体前34.61%,腹吸盘稍大于口吸盘,二吸盘横径之比为1.26±0.116:1,卵巢分4～6支,每支有2～3支指状分支。睾丸分4～5叶,中心块明显,每叶具指状或乳头状突起,排泄囊达肠支分叉处。体棘混生型。囊蚴呈微红色球状,大小为(525.16±38.61)×(500.19±31.7l)μm。与泰国标本相比,我国发现的哈氏并殖吸虫的成虫形态稍有变异。分子生物学初步研究表明,PCR扩增产物经BsaHI和StuI酶切(PCR-RFLP),琼脂糖凝胶电泳分离图谱完全一致。浙江宁海哈氏并殖吸虫ITS2基因有366碱基对,与泰国种群(AF159609)同源性为95.6%,CO1基因有390碱基对,与泰国种群(AF159600)同源性为89.7%。结论本研究从形态学和分子生物学进一步确认...

Objective To further identify Paragonimus harinasutai from Ninghai,Zhejiang,China.Methods 1.Morphology observation:The metacercaria of P.harinasutai were collected from Xixi village in Ninghai County,Zhejiang.The cats as animal model were infected for adult worms collection and morphology observation.2.Molecular biology study:The metacercariae were collected from Xixi villages in Ninghai County for ITS2 sequence analysis,CO1 sequence analysis and endonuclease BsaHI and StuI analysis(PCR-RFLP).Results The ra...