多重、非对称PCR-FP技术同步检测沙眼衣原体和解脲支原体

Detection of chlamydia trachomatis and ureaplasma urealyticum using fluorescence polarization based on a multiple and asymmetric PCR

目的:应用多重PCR-荧光偏振(FP)技术建立一种可靠、简单经济、快速的同步检测沙眼衣原体、解脲支原体的新方法,并推向临床应用.方法:以与沙眼衣原体dnaB样蛋白、解脲支原体UreC各血清型保守区序列互补的2对特异而无交叉反应的引物在同一反应体系中行多重、非对称聚合酶链反应扩增246例临床样本,应用沙眼衣原体、解脲支原体特异探针混合物与上述非对称扩增PCR产物孵育杂交,荧光偏振检测技术检测,并与常规检测方法测定结果比较.结果:应用多重、非对称PCR-FP技术同步检测泌尿生殖系感染临床标本246例,其中沙眼衣原体感染阳性56例,占22.7%;解脲支原体感染阳性62例,占25.2%;沙眼衣原体、解脲支原体混合感染阳性28例,占11.2%.与常规方法检测结果比较,两者对感染阳性的检测符合率为100%.结论:初步建立了具有良好的特异性、仅需一次扩增即可方便简单地检测沙眼衣原体、解脲支原体的同步检测新方法,对于相关感染的筛查及预防、预后判断具有重要价值.

AIM： To develop a simple, economical, accurate and practical method for detection of chlamydia trachomatis and ureaplasma urealyticum simultaneously by fluorescence polarization（FP） assay based on a multiple and asymmetric PCR. METHODS ： Two sets of primers were used to amplify chlamydia trachomatis and ureaplasma urealyticum DNA by a multiple and asymmetric PCR. The probes of chlamydia trachomatis and ureaplasma urealyticum labeled with different fluorophores were hybridized respectively with target amplicons. The infection was determined by the increased FP value. Two hundred and forty-six samples were subjected to common assay to evaluate the feasibility of this method. RESULTS： Of the 246 samples, 56 samples were identified as positive for chlamydia trachomatis, 62 samples were identified as positive for ureaplasma urealyticum and 28 samples were identified as positive for double infection by the multiple PCR-FP assay. The results obtained by the muhiple PCR-FP assay were all correct compared with the results by common assay. CONCLUSION： The proposed method allows a special, simple and economical detection for the simultaneous detection of chlamydia trachomatis and ureaplasma urealyticum by FP assay based on a multiple and asymmetric PCR.