16SrRNA、16S-23SrRNA基因测序分析检测主要血流感染病原菌比较

目的比较细菌16SrRNA、16S-23SrRNA基因测序分析在血流感染病原菌检测中的作用。方法提取临床上血流感染常见的金黄色葡萄菌、表皮葡萄球菌、大肠埃希菌、粪肠球菌、肺炎链球菌、铜绿假单胞菌、阴沟肠杆菌、鲍曼不动杆菌、洛菲不动杆菌、肺炎克雷伯杆菌、化脓性链球菌、奇异变形杆菌、潘尼变形杆菌、屎肠球菌、粘质沙雷菌、宋内志贺菌、产气肠杆菌、小肠结肠炎耶尔森菌、腐生葡萄球菌基因组DNA，运用16SrRNA、16S-23SrRNA基因进行PCR扩增。扩增产物经测序后在美国国家生物技术中心（NCBI）上进行比对分析，确定菌种。结果在所分析的19种临床血流感染常见细菌中，16SrRNA基因测序分析可将除粘质沙雷菌外的细菌鉴定到种的水平，但无法完全区分近缘种属；16S-23SrRNA成功鉴定17种细菌，除大肠埃希菌、宋内志贺菌外所有细菌均成功鉴定到单一种的水平。结论16S-23SrRNA基因可作为血流感染细菌检测较好的分子靶标。

Comparison of the role of 16S rRNA and 16S-23S rRNA gene sequence-based identification of bacteria in bloodsteam infection

Objective Comparison of the role of 16S rRNA gene and 16S-23S rRNA gene sequence- based identification of bacterial in bloodsteam infection. Methods Genomic DNA of common bacteria（Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Enterococcus ~aecalis, Streptococcus pneumoniae, Pseudomonas aeruginosa, Enterobacter cloacae, Acinetobacter baumannii, Acinetobacter lwoBTi, Klebsiella pneumoniae, Streptococcus pyogenes, Proteus mirabilis, Proteus penneri, Enterococcus ~aecium, Sen＇aria marcescens, Shigella sonnet~ Enterobacter aerogenes, Yersinia enterocolitica, Staphylococcus saprophyticus） in clinical bloodstream infection was extracted. Then, 16S rRNA gene and 16S-23 S rRNA gene were amplified by polymerase chain reaction（PCR）. The amplified products were analyzed in the national center biotechnology information（NCBI） of the United States in order to confirm the bacterial species. Results In 19 common bacteria of clinical bloodstream infection, 16S rRNA gene sequencing analysis could identified all bacteria to species level except Serratia marcescens but it couldn＇t distinguish close species; 16S-23S rRNA gene sequencing analysis identified 17 bacteria successfully and all bacteria except Escherichia coli and Shigella sonnei were identified to the single species level. Conclusion The 16S-23S rRNA gene can be used as a useful molecular target in the detection of bacteria in bloodstream infection.