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目的比较肺炎链球菌多重耐药株与敏感株标准参考菌之间的差异基因。方法使用抑制性消减杂交技术构建多重耐药肺炎链球菌差异DNA消减文库,经斑点杂交筛选阳性克隆后对部分DNA片段进行测序和同源分析。结果成功的构建了肺炎链球菌多重耐药株差异DNA消减文库,经斑点杂交的初步筛选,有17个仅能与多重耐药株DNA探针杂交,而不能与敏感株DNA探针杂交。对这17个克隆片段测序及基因库检索分析,发现2个未知新序列。结论从全基因角度研究肺炎链球菌多耐药菌株与标准菌株基因组DNA分子遗传差异,为发现、克隆肺炎链球菌多重耐药相关基因提供依据。
Objective To compare the differential genes between Streptococcus pneumoniae multi-drug resistant strains and reference strains of susceptible strains. Methods Suppression subtractive hybridization (PCR) was used to construct a subtractive cDNA library of multi-resistant Streptococcus pneumoniae. The positive clones were screened by dot blot hybridization and then sequenced and homologous analyzed. Results The DNA subtractive library of multi-drug resistant strains of Streptococcus pneumoniae was successfully constructed and 17 were screened by dot blot hybridization. Only 17 of them could only hybridize with multidrug resistant DNA probes and could not hybridize with sensitive DNA probes. The 17 cloned fragments were sequenced and analyzed by GenBank. Two unknown new sequences were found. Conclusion The molecular genetic differences between genotypes of Streptococcus pneumoniae multi-drug resistant strains and standard strains were studied from the perspective of whole genome, which provided the basis for discovering and cloning multi-drug resistant genes of Streptococcus pneumoniae.