【摘要】 目的制备卫氏并殖吸虫线粒体细胞色素C氧化酶亚单位Ⅰ(COⅠ)基因和核糖体DNA第二间隔区(ITS2)地高辛标记探针,探讨其对并殖吸虫的种别鉴定和诊断的作用。方法利用引物分别体外扩增获得卫氏并殖吸虫目的基因后凝胶电泳鉴定并且胶回收纯化上述两个基因片段。将酶切后的COⅠ基因和ITS2基因回收,地高辛标记成探针,尼龙膜上行斑点杂交观察。结果COⅠ探针有较好的特异性和敏感性。ITS2探针缺乏特异性。结论尼龙膜斑点杂交结果显示卫氏并殖吸虫的COⅠ基因片段可以作为种群鉴定的后备探针。更多还原

【Abstract】 Objective To prepare digoxigenin-Paragonimus westermani COⅠ and ITS2 genes labeled probe and evaluate for species identification. Methods The genes were labeled with digoxigenin and used as probes. DNA from the various parasites and rabbit control were applied to nylon transfer membranes,hybridized with labeled probes,and visualized according to the manufacturer’s instructions. Result The COⅠ probe hybridized well with DNA from Paragonimus westermani,but not with DNA from other species. In contrast,ITS2 could hybridize with DNA from both Paragonimu westermani and other species. Conclusion The COⅠ gene of the Paragonimus westermani labeled with digoxgenin probes can be used for species identification.更多还原