幽门螺杆菌16SrDNA指纹图和PCR-RFLP基因分型研究

Genotyping study on Helicobacter pylori by 16S rDNA fingerprinting and ureC PCR-RFLP patterns

目的 探讨我国幽门螺杆菌 (Helicobacterpylori,Hp)的基因多态性及其与相关疾病之间的关系。 方法 应用PCR掺入法制备 16SrRNA基因探针 ,建立 16SrDNA指纹图技术和尿素酶C(ureC)基因的PCR -RFLP法 ,并结合统计学软件进行聚类分析 ,对临床分离的 14株Hp进行基因分型。结果 14株Hp的 16SrDNA指纹图谱显示 10个HaeⅢ杂交带型和 12个HindⅢ杂交带型 ;ureC基因的变异度较小 ,14株Hp中有 12株PCR -RFLP图谱完全一致。通过杂交结果示意图和SPSS10 0软件进行聚类分析 ,HaeⅢ、HindⅢ及两种酶杂交结果的综合 ,均将 14株Hp分为两型。两种方法结果综合进行聚类分析 ,14株Hp在基因水平上分为三型。结论 差异显著的DNA指纹图谱说明了不同Hp菌株间基因组结构的高度变异性。 16SrDNA指纹图谱较ureC基因的PCR -RFLP谱型更敏感地表达Hp各菌株的变异 ,可准确有效地对Hp作出基因分型。

Aim To explore the genotypic diversity of H pylori and the relationship between the genotyping and diseases Methods 16S rRNA fingerprinting and PCR RFLP techniques were used to classify 14 clinical isolates of H pylori Results 14 isolates of H pylori were devived into 10 hybridization patterns with HaeⅢ and 12 hybridization patterns with HindⅢ The diversity of ureC gene in this study was less When both of the results were combined,14 isolates of H pylori were classfied into three types by using cluster analysis of SPSS 10 0 Software Conclusion H pylori were heterogeneous and represented a wide diversity of genetic pattern 16S rDNA fingerprinting and PCR RFLP of ureC could be a reliable,highly discriminatory and reproducibility typing scheme 16s rDNA fingerprinting was better than PCR RFLP of ureC for H pylori typing The result couldn't reflect the relationship between different genotypes and different diseases