论文部分内容阅读
目的探讨巢式聚合酶链反应(PCR)方法在快速检测新生儿败血症病原菌中的应用价值。方法以细菌16S rRNA基因为扩增靶序列,设计一对所有细菌通用引物和一对革兰阴性菌特有引物,用巢式PCR方法扩增已知病原菌,检测其特异性。用巢式PCR方法和培养法同时检测新生儿败血症血液,对2种方法的检测结果进行比较分析。结果已知实验菌株经通用引物扩增后均获得920 bp产物,革兰阴性菌经革兰阴性菌特有引物扩增后获得353 bp产物,而革兰阳性菌、人类基因组DNA、巨细胞病毒和白色假丝酵母菌无相对应产物。2种方法检测病原菌革兰分型吻合度一致,但巢式PCR方法阳性率高。结论巢式PCR方法检测新生儿败血症病原菌,具有快速、敏感度高等优点,并能对病原菌进行革兰分型,具有一定的实用价值。
Objective To explore the value of nested polymerase chain reaction (PCR) in the rapid detection of neonatal sepsis pathogens. Methods A pair of bacterial universal primers and a pair of primers specific to Gram - negative bacteria were designed by using bacterial 16S rRNA gene as target sequence. Known pathogenic bacteria were amplified by nested PCR and their specificity was tested. Nested PCR and culture were used to detect the blood of neonatal sepsis at the same time. The results of two methods were compared and analyzed. Results It was known that the amplified products of the experimental strains obtained 920 bp products after amplification by universal primers. The products of 353 bp were obtained after the Gram-negative bacteria were amplified by the primers specific to Gram-negative bacteria. The Gram-positive bacteria, human genomic DNA, cytomegalovirus and Candida albicans no corresponding product. There was no difference between the two methods in detecting gram-positive of pathogenic bacteria, but the positive rate of nested PCR was high. Conclusion The nested PCR method for detection of neonatal septicemia pathogens, with fast, high sensitivity, and can Gram-type pathogens, has a certain practical value.