16S rRNA基因在快速诊断新生儿败血症的病原菌研究

Research on a rapid method for the detection of bacteia in neonatal septicemia by using 16S rRNA gene

目的探讨血液16S rRNA基因检测在新生儿败血症诊断中的应用价值。方法分析细菌16S rRNA基因保守区,设计一对通用引物扩增已知实验菌株,检测其特异性,用倍比稀释法检测其敏感性,同时进行血培养。结果已知实验菌株均获得920bp扩增产物,对照组中的人类基因组DNA、HBV-DNA和白色假丝酵母菌无相应产物。敏感性测试能达到1pg大肠杆菌DNA。PCR阳性率为31.7%(20/63),血培养阳性率为14.3%(9/63),两者比较有显著性差异(P<0.05)。结论 PCR检测血液细菌16S rRNA基因,具有特异性强,敏感度高等特点,能在临床推广应用。

Objective： To explore the significance of 16S rRNA gene in diagnosis of neonatal septicemia. Methods： We analysed bacterial 16S rRNA genes in conservative districts and designed a pair of universal primers for amplification of a known experimental strains for specific detection, with double dilution assay for detection of sensitivity, and blood culture was conducted simultaneously. Results ： The known experimental strains were amplified and products were 920bp, but human genome DNA, HBV - DNA and candi- da albicans showed no corresponding products. Sensitivity test showed that it could detect as low as lpg of E. coli DNA. The positive rate of PCR was 31.7% （20/63）, the positive rate of blood culture was 14. 3% （9/63）, more significant differences between the two group （P 〈 0. 05 ）. Conclusion ： The technology of PCR has strong specificity and high sensitivity, it can be used for clinical.