16SrRNA基因序列分析用于诊断病原性细菌感染初步研究

目的建立一种可适用于多种病原菌检测的方法,以提高病原学诊断率,有利于传染病的病原学监测。方法细菌16SrRNA基因通用引物对直接从血液、尿液、脑脊液标本中提取的细菌DNA进行PCR扩增,并对16SrRNA基因扩增产物进行克隆和序列分析。结果检测了28份临床标本,16SrRNA基因序列分析与尿液细菌培养结果不完全吻合;血液和脑脊液的16SrRNA的检出率均高于细菌培养。结论16SrRNA基因检测分析和细菌培养均具有良好的诊断效果,而针对16SrRNA基因的PCR直接扩增核酸并进行序列分析可对分离培养阴性的标本进行有效的补充诊断,可检测到多种病原菌如肠球菌,大肠杆菌,表皮葡萄球菌,牛链球菌,猪链球菌,布鲁氏菌,表皮葡萄球菌和脑膜炎球菌等,为疾病的诊断提供良好的线索。

Gene sequence analysis of 16S rRNA for the use of diagnosis in primary bacterial infections

To search for a laboratory method suitable for use in the diagnosis of multiple pathogens in order to improve the efficiency of etiological diagnosis and surveillance of infectious diseases, the 16S rRNA gene sequence of bacterial DNA taken from blood, urine and CSF specimens was amplified with universal primers, and the amplified products were then cloned and sequenced.Among the 28 clinical specimens examined, the results of the 16S rRNA gene sequence analysis did not completely correlate with the results obtained from the bacterial cultures of urine, but the detection rates of 16S rRNA gene sequence analysis for bacteria in blood and CSF were significantly higher than those obtained by bacterial cultures.Different kinds of bacteria, such as Enterococcus, E.coli, Staphylococcus epidermidis, Streptococcus suis, Styreptococcus bovis, Brucella species and Neisseria meningitidis can be detected by both the 16S rRNA gene sequence analysis and bacterial culturing, but the former may be used as an additional approach for diagnosis, especially when the isolation cultivation method shows a negative result.