肝内胆管结石和胆汁中细菌多样性分析

Analysis of bacterial diversity in human hepatolithiasis and bile

运用培养法和聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术对6个病人肝内胆管结石和胆汁中的细菌群落进行了分析.胰蛋白胨大豆肉汤(TSB)培养法结果显示,从6个胆石和5个胆汁样品中分别分离到15株和12株细菌,均属于Escherichia和Enterococcus两个属,而同一病人胆石及胆汁中细菌种类并不完全相同,其中一份胆汁样在两种方法检测中皆为阴性;PCR-DGGE结果表明,胆石及胆汁样品中除培养法中发现的两个属外,还包含Pseudomonas、Morganella、Citrobacter、Baterioides、Serratia、Clostridiales和Aeromonas共7个属的细菌.在5组PCR阳性的胆石和胆汁的DGGE指纹图谱相似性分析中,每组内胆石和胆汁间细菌群落结构的最高相似性仅为32.3%.两种检测方法的结果表明,Escherichia coli在胆石和胆汁中的检出率最高,胆石和胆汁中细菌多样性存在差异,PCRDGGE技术比培养法能更好检测到胆石和胆汁中的细菌多样性.

The bacterial community in human hepatolithiasis and bile coming from six patients were examined by culture-dependent approach and polymerase chain reaction-denaturing gradient gel electropho- resis （PCR-DGGE）. A total of 15 and 12 bacterial strains were isolated from hepatolithiasis and bile samples respectively by culturing in tryptic soy broth （TSB） and further identified as Escherichia or En- terococcus spp, but the bacteria community may not necessarily be the same in hepatolithiasis and corre- sponding bile from one patient. PCR-DGGE showed that, besides Escherichia and Enterococcus, other 7 genera of Pseudomonas, Morganella, Citrobacter, Baterioides, Serratia, Clostridiales and Aeromonas were disclosed The highest similarity coefficient between hepatolithiasis and bile was only 32. 2% by similarity analysis in 5 patients. By these two detection approaches, Escherichia coli has the highest detection rate, and bacterial diversity of hepatolithlasis find bile was different. Compared with culture-dependent approach, more diversi{ied bacterial community in both human hepatolithiasis and bile can be observed by PCR-DGGE.