PCR法检测细菌23S rDNA在感染性疾病快速诊断中的应用研究  

Applied Investigation on PCR Method of Amplifying Bacterial 23S Ribosomal DNA in the Rapid Diagnosis of Infectious Diseases

作　　者：阮秋蓉[1] 李国军[2] 王卫华[2] 李洪涛[2] 覃惠敏[2] 宋建新[2] 

机构地区：[1]华中科技大学同济医学院基础医学院病理学系,武汉430030 [2]华中科技大学同济医学院附属同济医院感染科

出　　处：《华中科技大学学报（医学版）》2004年第3期273-275,共3页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：湖北省科技攻关计划项目 (No 2 0 0 3AA30 1C6 1)

摘　　要：目的　研究建立快速检测细菌DNA的方法。方法　设计一对共同引物 ,应用聚合酶链反应 (PCR)法 ,扩增细菌核糖体 2 3S亚单位基因 2 3SrDNA。对 2 3种临床常见致病菌、培养物及临床病例标本采用该方法进行检测 ,并与常规细菌培养法比较。结果　该方法可检测细菌下限为 2 5CFU ,与真菌、乙肝病毒等无交叉反应。对纯菌及培养物均可检出一条明显DNA条带 ,革兰阴性细菌约 35 0bp ,革兰阳性细菌约 4 0 0bp。临床标本 2 3SrDNA检出率明显高于常规培养方法 (P <0 0 1)。结论　应用所设计的共同引物 ,检测细菌 2 3SrDNA ,敏感性和特异性好 ,比常规培养法快速、准确。Objective To establish a rapid method of detecting bacterial 23S ribosomal DNA (23S rDNA). Methods A pair of universal primers were designed to amplify bacterial 23S rDNA by PCR. Compared with routine bacterial culture method, this novel PCR method was used to detect 23S rDNA of 23 kinds of pure bacteria, culture bottles and clinical specimens. Results This method could detect bacteria to the lowest concentration of 2.5 CFU, and had no cross reaction with human genome and hepatitis B virus DNA. A band of approximately 350 bp was produced with gram-negative bacteria, and a band of 400 bp with gram-positive bacteria in pure bacteria culture and culture bottles. It showed higher positive rate in detecting clinical specimens than routine bacterial culture (P<0.01). Conclusion This PCR method with universal primers shows higher sensitivity and specificity in detecting bacterial 23S rDNA and is more rapid and accurate than routine culture method.

关 键 词：细菌培养法 DNA 感染性疾病 PCR法 快速诊断 常见致病菌 革兰阳性细菌 核糖体 真菌 引物 

分 类 号：R446[医药卫生—诊断学] R378[医药卫生—临床医学]