【摘要】 目的初步建立PCR技术直接快速检测血培养阳性瓶中大肠埃希菌的方法。方法采用美国BiOsticBacteremia DNA Isolation Kit(血培养液DNA提取试剂盒),提取102份经革兰染色确定为革兰阴性杆菌的血培养阳性标本细菌DNA,用自行设计的大肠埃希菌特异性引物进行PCR扩增,扩增产物经琼脂糖凝胶电泳进行细菌鉴定,结果与VITEK-Ⅱ微生物全自动分析仪鉴定结果进行比较。结果 102株革兰阴性杆菌经VITEK-Ⅱ微生物全自动分析仪鉴定出33株大肠埃希菌,PCR方法鉴定出31株,两者进行配对χ2检验,P>0.01,两种检测方法无差别。结论该PCR技术可用于快速检测血培养标本中大肠埃希菌,耗时3~4小时;该技术也可为其他病原菌的PCR检测方法的建立奠定基础。更多还原

【Abstract】 Objective To establish a polymerize chain reaction method for direct detection of E.coli in positive blood culture bottles. Method American BiOstic Bacteremia DNA Isolation Kit(blood cultures of DNA extraction kit) was used to extract bacterial genomic DNA from 102 positive blood culture specimens of Gram-negative bacilli determined by the Gram staining. Self-designed E.coli specific primers was used for PCR amplification and the identification of bacteria to the amplification products was performed by agarose gel electrophoresis. The result was compared with that of VITEK- Ⅱ automatic analyzer identification. Result 33 strains and 31 strains of E.coli were detected in 102 strains of Gramnegative bacilli by VITEK- Ⅱ microbial automatic analyzer and polymerase chain reaction(PCR) respectively.χ2 matching test indicated that there was no statistical difference between the two kinds of detection methods(P > 0.01). Conclusion The PCR method can be applied for the rapid detection of E.coli in blood cultures, which takes 3 ~ 4 hours. It also builds a foundation for PCR detection methods of other pathogens.更多还原