鹦鹉热嗜衣原体两对荧光定量PCR引物的应用与比较

Comparison on two pairs of primer used in fluorescent quantitative assay to detect genes encoding the major outer membrane protein and the polymorphic membrane protein of Chlamydophila psittaci

针对鹦鹉热嗜衣原体(Cps)的主要外膜蛋白(MOMP)基因和多形态膜蛋白(PMP)基因分别设计引物,建立荧光定量PCR方法,比较两对引物的灵敏度和特异性。试验结果表明,两对引物均可用于Cps检测,在样本中靶标浓度高时,PMP引物的检测灵敏度优于MOMP引物;但前者的扩增效率低于后者,后者更适合用于Cps的实时定量PCR检测。相对定量研究表明国内不同Cps流行株的PMP基因在基因组上的拷贝数可能存在较大差异。

Two pairs of primer targeting to the genes encoding the major outer membrane protein （MOMP） and the polymorphic membrane protein （PMP） of Chlamydophila psittaci （Cps） designed and used in the fluorescent quantitative PCR assay in the detection of this microorganism were compared in the their sensitivity and specificity. It was demonstrated that both pairs of primer could be used successfully to detect the presence of Cps of the epidemic strains isolated in China. The specificity of PMP primer used was higher than that of MOMP primer. In case of high target concentration in samples, the sensitivity of the former was also superior to that of the latter, but the amplification efficiency of the former was lower than that of the latter. The MOMP primer seemed to be more suitable to detect Cps by using the real-time quantitative assay. As demonstrated by the real-time quantitative PCR assay, great difference in the genomic copy numbers of the PMP gene existed in the epidemic strains of Cps isolated in China.