Bacterial and Enteric Diseases Program, National Microbiology Laboratory, Winnipeg, Manitoba.
Can J Infect Dis Med Microbiol. 2005 Jan;16(1):15-25. doi: 10.1155/2005/323082.
The present article describes the laboratory diagnosis of Neisseria gonorrhoeae by culturing of the organism from different types of clinical specimens followed by confirmatory tests. The success of culture methods requires good quality collection and transport of clinical specimens. The present guide describes the media requirements and cultural conditions for N gonorrhoeae growth and the characteristics for a presumptive identification of N gonorrhoeae. Confirmatory tests include biochemical tests, chromogenic enzyme substrate tests, immunoassays and nucleic acid methods. Nucleic acid detection methods include either amplification-based methods or nonamplification tests, and are increasingly used in clinical laboratories where a viable culture is not possible to obtain. Nucleic acid methods can also be used to detect the presence of low numbers in a specimen. Nucleic acid detection methods need confirmation with another amplification method or gene target. Controls must be included to ensure true positive and negative results, and to rule out nucleic acid contamination. Monitoring of antimicrobial susceptibilities of N gonorrhoeae is important to investigate treatment failure and to evaluate the efficacy of currently recommended therapies. Many methods for the characterization of N gonorrhoeae require cultures. The useful typing methods for determining strain relatedness include auxotyping, serotyping, plasmid profile analysis, DNA sequencing of the porB gene and pulsed-field gel electrophoresis. Quality assurance programs for diagnostic testing and antimicrobial susceptibility testing is reviewed.
本文描述了通过培养来自不同类型临床标本的生物体并进行确证试验来对淋病奈瑟菌进行实验室诊断。培养方法的成功需要良好质量的临床标本采集和运输。本指南描述了淋病奈瑟菌生长的培养基要求和培养条件,以及淋病奈瑟菌的推定鉴定特征。确证试验包括生化试验、显色酶底物试验、免疫测定和核酸方法。核酸检测方法包括基于扩增的方法或非扩增试验,并且越来越多地用于临床实验室,在这些实验室中不可能获得可行的培养物。核酸方法也可用于检测标本中低数量的存在。核酸检测方法需要用另一种扩增方法或基因靶标进行确认。必须包含对照以确保真实的阳性和阴性结果,并排除核酸污染。监测淋病奈瑟菌的抗菌药物敏感性对于研究治疗失败和评估目前推荐的治疗方法的疗效非常重要。许多用于淋病奈瑟菌特征描述的方法都需要培养。用于确定菌株相关性的有用分型方法包括辅助表型分析、血清分型、质粒图谱分析、porB 基因的 DNA 测序和脉冲场凝胶电泳。对诊断检测和抗菌药物敏感性检测的质量保证计划进行了审查。