用随机引物多态性DNA聚合酶链反应鉴别新的裂体吸虫X  

IDENTIFICATION OF SCHISTOSOMA X WITH RAPD-PCR

作　　者：徐国余[1] 陈广梅[1] 田海生[2] 朱昌亮[2] 

机构地区：[1]江苏省南京市疾病预防控制中心,南京210003 [2]南京医科大学病原生物学与免疫学系

出　　处：《中国血吸虫病防治杂志》2003年第1期39-40,共2页Chinese Journal of Schistosomiasis Control

摘　　要：目的　鉴定新的裂体吸虫χ的基因组 DNA。方法　应用随机引物多态性 DNA聚合酶链反应 (RAPD- PCR)对裂体吸虫χ(S.χ)和日本血吸虫 (S.j)的基因组 DNA进行分析鉴别。感染了S.χ和 S.j尾蚴的兔 4 5 d后杀死 ,各取成虫 5 0条 ,磨碎 ,用苯酚—氯仿抽提法提取基因组 DNA。反应在 PCR扩增仪上进行 ,应用 2 9个随机引物 ,1.4 %琼脂糖凝胶电泳后观察、照相。结果　 2 9个引物中 2个引物 ,J0 1 碱基 CCCGGCATAA和 L1 2 碱基 GGGCGGTACT的扩增产物在两者间显示出明显的差异带。引物 J0 1 S.χ特异性条带 1条 ,S.j特异性条带 1条。引物 L1 2 S.χ特异性条带 3条 ,S.j特异性条带 1条。结论　 S.χ和 S.j的基因组 DNA在 J0 1 和 L1 2 的 6个位点的序列有所不同 ,出现了差异带 ,这在种间进化方面有鉴别价值。Objective To identify genomic DNA of Schistosoma χ. Methods Amplification of genomic DNA by the random-amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) with 10-base pair was used. The 50 worms were collected from rabbits infected with cercariae of S.χ and Schistosoma japonicum(S.j.) respectively. RAPD-PCR were performed on PCR-2400 according to the manufature's instruction. And 29 primers were adopted from Operon Company. The samples were run on 1.4% sepharose. Results RAPD fragments produced were various in quantity(4-12 bands)and size(0.5-5.2Kb) in S.χ and S.j. , most of about 224 bands produced with 27 different primers were common, but 6 differential bands produced with 2 primers (J 01 CCCGGCATAA and L 12 GGGCGGTACT) of the 29 primers were found, 4 and 2 of the 6 differential bands seen in the S.χ and S.j. respectively. Conclusion These specific fragments found in S.χ and S.j. may be used as molecular markers for the identification of S.χ and S.j.

关 键 词：裂体吸虫x DNA 多态性 聚合酶链反应 日本血吸虫 

分 类 号：R383.24[医药卫生—医学寄生虫学]