16SrRNA-PCR在淋球菌引起的泌尿生殖道感染检测中的应用  

Utilization of 16SrRNA-PCR in Detection of Genitourinary Tract Infection Caused by Neisseria Gonorrhoeae

作　　者：谢良伊[1] 蔡瑞云[1] 

机构地区：[1]湖南省人民医院检验科,湖南长沙410005

出　　处：《实用预防医学》2010年第3期571-573,共3页Practical Preventive Medicine

摘　　要：目的建立一种快速、特异性、敏感地检测淋球菌致泌尿生殖道感染的16SrRNA-PCR基因诊断方法,以了解泌尿生殖道感染中淋球菌的感染情况。方法以淋球菌16S核糖体rRNA基因为扩增靶点,设计一对特异性的引物扩增靶基因片段,扩增的片段长度为260bp,扩增产物通过ABI3130 DNA Analyzer进行测序,测序结果同Genebank报道的序列进行比较,评价该方法的特异性;对淋球菌模板进行10倍比稀释,进行16SrRNA-PCR,对其敏感性进行分析,对PCR扩增产物进行电泳和测序鉴定。结果泌尿生殖道感染中通过16SrRNA-PCR扩增可检出淋球菌16SrRNA基因,通过序列分析证实了PCR产物的特异性;在敏感性检测中16SrRNA-PCR检测淋球菌模板的下限为6.41×10-4μg/ml。结论本研究建立的16SrRNA-PCR法检测泌尿生殖道淋球菌感染具有简便、特异、快速的特点,可作为临床上女性宫颈分泌物及尿液标本的检测。Objective To establish a rapid,specific and sensitive polymerase chain reaction （16SrRNA-PCR） assay for the detection of genitourinary tract infection caused by Neisseria gonorrhoeae,and to know the prevalence of Neisseria gonorrhoeae in genitourinary tract infection. Methods According to the sequences of Neisseria gonorrhoeae 16SrRNA,a couple of specific primers were designed to amplify the target gene sequence,and the size of the amplified fragment was 260 bp. The amplified products were analyzed by ABI3130 DNA Analyzer,the results of sequence analysis were compared with the sequence report of Genebank,and then the specificity of the sequence analysis method was evaluated. The template was serially diluted by 10 times and then amplified it by 16SrRNA-PCR to verify the sensitivity and the detection limitation,at last the PCR products were assayed by electrophoresis and DNA sequence analysis. Results We could detect Neisseria gonorrhoeae through 16SrRNA PCR in genitourinary tract infection,the specificity was verified by gene analysis and the lowest limit we had detected in the sensibility test was 6.41×10-4 ug/ml. Conclusions The 16SrRNA-PCR assay is a convenient,specific and sensitive method in detecting Neisseria gonorrhoeae infection,and it can be used for the clinical detection of cervical secretion and urine sample.

关 键 词：16SrRNA-PCR 淋球菌感染 基因测序 

分 类 号：R378.16[医药卫生—病原生物学]