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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 investigate the value of blood 16S rRNA gene detection in the diagnosis of neonatal sepsis. Methods The conserved region of bacterial 16S rRNA gene was analyzed. A pair of universal primers was designed to amplify the known experimental strains. The specificity of the 16S rRNA gene was tested. The sensitivity of the 16S rRNA gene was tested by multiplicity dilution assay and the blood cultures were also performed. Results It is known that all the experimental strains obtained the 920 bp amplification product. There was no corresponding product of the human genomic DNA, HBV-DNA and Candida albicans in the control group. Sensitivity test can reach 1pg E. coli DNA. The positive rate of PCR was 31.7% (20/63), and the positive rate of blood culture was 14.3% (9/63). There was a significant difference between them (P <0.05). Conclusion PCR detection of blood bacterial 16S rRNA gene has the characteristics of strong specificity and high sensitivity, which can be widely used in clinic.