摘要
目的探讨双重荧光定量PCR技术的优化条件,建立基于TaqMan探针技术荧光定量法检测同时检测解脲支原体和巨细胞病毒的新方法。方法分别采用普通定性PCR扩增母婴垂直传播常见的病原体(解脲支原体和巨细胞病毒)测序鉴定,然后分别采用TaqMan探针的单重和双重定量PCR技术对解脲支原体和巨细胞病毒同时定性定量检测。结果解脲支原体和巨细胞病毒单种定性PCR检测均为阳性,TaqMan探针单重和双重定量PCR检测解脲支原体和巨细胞病毒阳性率和特异性均为100%,相同样品TaqMan探针单重、双重定量PCR分别检测的结果符合率100%。结论 TaqMan探针双重荧光定量PCR技术可同时检测两种靶分子,结果可靠,应用前景广阔。
Objective To optimize the condition of Duplex fluorescence quantitative PCR and establish a new assay for Ureaplasma urealytieum （ UU ） and Cytomegalovirus （ CMV ） detection based on TaqMan duplex fluorescence quantitative PCR. Methods UU and CMV were identified by sequence analysing via qualitative PCR and then quantitatively detected by TaqMan single and duplex fluorescence quantitative PCR respectively. Results Single qualitative PCR gave positive results in the detection of UU and CMV. The positive rates and specificity of both TaqMan single and duplex fluorescence quantitative PCR in detecting UU and CMV achieved 100%. UU and CMV were simultaneously detected by TaqMan single and duplex fluorescence quantitative PCR. Conclusion TaqMan duplex fluorescence quantitative PCR assay can simultaneously detect Ureaplasma urealyticum and Cytomegalovirus with reliable results, thus has a board prospect of application.
出处
《中国微生态学杂志》
CAS
CSCD
2014年第2期214-216,220,共4页
Chinese Journal of Microecology
基金
杭州市科技局社会发展科研计划资助项目(20130733Q04)
杭州市卫生科技计划重大项目(2012ZD001)
