16SrRNA基因检测在自发性细菏性腹膜炎早期诊断中的应用

目的探讨16SrRNA基因检测在快速诊断自发性细菌性腹膜炎（SBP）中的临床应用价值。方法通过对细菌16SrRNA基因保守区和变异区的序列分析，设计通用引物，对已知实验室病原菌、人类基因组DNA、巨细胞病毒和白色假丝酵母菌进行聚合酶链式反应（PCR）扩增，检测方法的特异性；采用倍比稀释法进行方法敏感性检测。对SBP患者的腹水同时进行PCR扩增和普通培养，比较两种方法的阳性率。结果已知茵株均获得920bp扩增产物，人类基因组DNA、巨细胞病毒和白色假丝酵母茵无相应产物，敏感性能达到lpg大肠埃希茵DNA。PCR法阳性率显著高于培养法（P〈0．05）。结论PcR技术检测腹水病原菌16SrRNA基因，具有特异性强，敏感度高等特点，在早期、快速诊断SBP方面具有重要临床应用价值。

Objective To explore the clinical application value of detection 16S rRNA gene in the rapid diagnosis of spontaneous bacterial peritonitis （ SBP ） . Methods Universal primers were designed through a known pathogen of a conservative district and variation of 16S rRNA gene sequence analysis. The specificity was detected through PCR amplifying known pathogen, human genomic DNA, cytomegalovirus and Candida albicans.The sensitivity was detected through doubling dilution. The ascites were done by PCR Method and culture Method at the same time, the positive rate was compared by the two Method s. Results The known pathogen were amplified and products were 920bp, but human genomic DNA, cytomegalovirus and Candida albicans showed no corresponding products. Sensitivity showed that it could detect as low as 1 pg of Escherichia coli. DNA.The positive rate of PCR Method was significantly higher than that of culture Method （ P〈0.05 ） . Conclusion The technology of PCR has strong specificity and high sensitivity, it has important clinical application value in early and rapid diagnosis SBP.