摘要
为快速准确鉴定诺卡菌,首先设计针对诺卡菌rpoB、secA1、16S rRNA基因的引物,利用单重聚合酶链反应(PCR)和测序验证引物的特异度,建立多重PCR鉴定系统,在同一反应体系和条件下对44株诺卡菌标准株、44株临床分离株和7株对照株进行扩增。结果显示,利用单对引物对其中2株诺卡菌(标准株DSM43003、临床株CDC 51)进行扩增,出现的条带均为与目的片段长度一致的单一条带,经测序并经基本局部比对搜索工具(BLAST)验证扩增片段为目的基因。建立的多重PCR结果显示,44株诺卡菌标准株中有43株(97.7%)、44株临床分离株中有42株(95.5%)rpoB、secA1、16S rRNA这3条片段均显示,7株对照株均未显示条带。结果提示,本研究建立的多重PCR简单、快速、灵敏度高、特异度好,适用于诺卡菌的快速鉴定。
In order to identify Nocardia quickly and accurately , three polymerase chain reaction （PCR ） methods were designed for the detection of rpoB , secA1 and 16 S rRNA genes respectively .The specificity of each PCR-based detection was verified .Then multiple PCR method was established ,and the sensitivity and specificity were tested by 44 Nocardia standard strains ,44 clinical isolates and 7 reference strains under the same conditions .The results demonstrated that 2 strains of Nocardia （DSM 43003 and CDC 51） were positive for the PCR reactions .The PCR products were verified by sequencing and Basic Local Alignment Search Tool （BLAST） .With the established multiple PCR method ,rpoB ,secA1 and 16S rRNA segments were detected in 43 of 44 （97 .7% ） Nocardia standard strains and 42 of 44 clinical isolate （95 .5% ） .There were no bands obtained in 7 reference strains .The specificity met the test requirement .It is concluded that multiple PCR method is fast , accurate , specific and sensitive . It can be used for identifying Nocardia strains .
出处
《微生物与感染》
2014年第2期107-111,共5页
Journal of Microbes and Infections
基金
"十二五"国家科技重大专项(2012ZX10004401
2013ZX10004221
2013ZX10004218)
公益性卫生行业科研专项(201302006)
关键词
诺卡菌
多重聚合酶链反应
鉴定
Nocardia
Multiple polymerase chain reaction
Identification
作者简介
通信作者：杨海燕E-mail：yhy@zzu．edu．cn
通信作者：李振军E-mail：1izhenjun@icdc．cn
