随机扩增多态性DNA在淋球菌基因分型中的应用

Application of random amplification polymorphic DNA in the genotyping of Neisseria gonorrhoeae

目的 建立随机扩增多态性DNA(RAPD)对淋球菌进行基因分型的方法。方法 用4种不同的预处理方法提取淋球菌基因组DNA,对其优劣进行比较;并运用RAPD对淋球菌菌株进行区分及对经性伴传播的淋病病例进行RAPD指纹图谱比较。结果 运用经典的十六烷基三乙基溴化铵法可以抽提较完整的基因组DNA,获得良好的RAPD指纹图谱;各菌株的RAPD指纹图谱间有明显不同DNA多态性;性伴传播的病例中获得了非常相似的RAPD指纹。结论 选择最佳的DNA抽提技术,运用RAPD可以对淋球菌进行有效基因分型,并可用于分子流行病学对传染源的追踪。

Objective To set up random amplified polymorphic DNAs (RAPD) method in genotyping Neisseria gonorrhoeae on DNA level, and to explore its use to trace the source of infection. Methods Four different pretreatments were used to extract the Neisseria gonorrhoeae genomic DNA with its advantages and disadvantages compared. Arbitrary sequence was then used to ampify the genomic DNA of Neisseria gonorrhoeae and RAPD fingerprint maps was applied to distinct the Neisseria gonorrhoeae strains Finally, RAPD fingerprint of Neisseria gonorrhoeae strain between paitient and his/her sexual partner was compared. Results Cetyltrimethylammonium bromide method was classical in extracting genomic DNA, and could get integrated genomic DNA and good fingerprint maps, since main segments were common to all the Neisseria gonorrhoeae but some were different among strains so that the frinerprint of different Neisseria gonorrhoeae were distinctive. However, fingerprint maps of Neisseria gonorrhoeae collected from sex partners were quite similar. Conclusion Based on genomic levels, effective fingerprint maps could be identified and to classify the Neisseria gonorrhoeae into different genotypes. RAPD fingerprint maps could be used to trace the source of infection.