23S rRNA基因在临床常见致病菌检测中的应用

目的　根据临床常见致病菌 2 3SrRNA基因序列的差异 ,建立可初步鉴别革兰阴性菌和革兰阳性菌的分子生物学方法。方法　分析常见细菌的 2 3SrRNA基因序列 ,设计相应引物。采用多重PCR扩增标准菌株及临床分离株 2 3SrRNA基因 ,并根据电泳结果作出初步分类。结果　革兰阴性菌株均出现 1条DNA电泳条带(约为 35 0bp) ,而革兰阳性菌株则出现 2条电泳条带 (约为 2 5 0和 4 0 0bp)。 6 0株临床分离菌经PCR扩增、电泳后 ,电泳分析结果与常规鉴定结果符合率达 10 0 %。结论　 2 3SrRNA基因检测用于细菌初步分类鉴定 ,具有快速、灵敏、准确的特点 ,可为细菌感染的实验室诊断提供客观依据

Application of 23S rRNA gene for the detection of common bacteria

Objective To analyze the 23S rRNA gene sequences of common bacteria, and establish a molecular biological technique to detect and differentiate Gram positive and Gram negative bacteria based on them. Methods After analyzing the 23S rRNA gene sequences of 6 standard strains, primers were designed. Standard strains and clinical isolates were amplified by a multiplex PCR. These bacteria were detected and differentiated according to the result of gel electrophoresis. Results Gram negative strains presented one band (about 350 bp) and Gram positive strains presented two bands (about 250 and 400 bp). 60 clinical isolates were identified with this method, and the coincidence rate, compared with the results of routine assay, was 100%. Conclusions The results indicate that using 23S rRNA gene to detect and differentiate bacteria is a specific, sensitive and rapid technique for laboratory diagnosis of bacteria infection.