论文部分内容阅读
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 investigate the optimization conditions of dual-fluorescence quantitative PCR and to establish a new fluorescence quantitative detection method for detection of Ureaplasma urealyticum and cytomegalovirus based on TaqMan probe technology. Methods The common pathogens (Ureaplasma urealyticum and Cytomegalovirus) of vertical transmission in mother and infant were amplified by general qualitative PCR respectively. Sequencing was performed by using single and double quantitative PCR with TaqMan probe. The results showed that Ureaplasma urealyticum and cytomegalovirus simultaneously Qualitative and quantitative testing. Results The results of single qualitative PCR detection of Ureaplasma urealyticum and cytomegalovirus were all positive. The positive rates and specificities of Ureaplasma urealyticum and cytomegalovirus were all 100% detected by TaqMan probe single and double quantitative PCR. The same sample TaqMan probe single Heavy and double quantitative PCR test results were 100%. Conclusion The TaqMan probe dual fluorescence quantitative PCR technology can detect two target molecules simultaneously, with reliable results and broad application prospects.