摘要
目的建立咽拭子标本中EV71和Cox A16型肠道病毒逆转录聚合酶链反应(RT-PCR)检测方法。方法根据GENBANK数据库中EV71和Cox A16型肠道病毒的基因序列,利用CLUSTAL软件进行序列比对分析,选择基因组的保守序列,并借助引物设计生物信息学软件,设计筛选出一套针对EV71和Cox A16型肠道病毒的RT-PCR扩增的引物。从特异性、最低检测限、重复性等方面进行评估,建立了EV71和Cox A16型肠道病毒RT-PCR检测方法。结果RT-PCR方法对EV71和COXA16病毒的最低检测量分别为63 TCID50/ml和32 TCID50/ml。其他病原微生物及人类基因组在该系统无反应信号,显示方法特异性良好。结论RT-PCR对手足口病病原体检测准确性高、特异性强,具有快速、经济等特点,适合手足口病的早期诊断。
Objective To study a RT-PCR method for detecting EV71 and Cox A16 virus from throat swab specimen. Methods Specific primers were designed based on highly conserved region of EV71 and Cox A16 depending on bioinformatics software. Specificity, minimum detection limits and repeatability of the method were assessed. Results The minimum detection limits of the method for EV71 and Cox A16 virus were 63 TCIDsdml and 32 TCIDsdml, respectively. Other pathogenic organisms and human genome in the method didn＇t show any response signal, displaying good specificity of the method. Conclusion Our results indicate RT-PCR offers a rapid, sensitive, specific and cheap method to detect pathogen of hand-food and mouth disease.
出处
《中国国境卫生检疫杂志》
CAS
2012年第4期228-231,共4页
Chinese Journal of Frontier Health and Quarantine
基金
深圳检验检疫局科研基金项目(SZ2009004)
