鹦鹉热嗜衣原体实时定量PCR检测方法的建立

Establishment of real-time quantitative PCR for detection of Chlamydophila psittaci

目的建立一种简便、快速、特异的荧光定量PCR检测方法,用于鹦鹉热嗜衣原体的快速检测。方法以主要外膜蛋白(MOMP)基因为靶序列设计特异性引物,采用SYBR GreenⅠ随机渗入法建立实时定量PCR检测方法。结果循环阈值(Ct)与标准DNA模板在1.0×102-1.0×107拷贝/μl浓度范围内呈良好的线性关系,相关系数为0.989。该方法用于鹦鹉热嗜衣原体的检测具有很高的特异性,其敏感性与常规PCR相比可以提高100倍。结论本研究建立的检测鹦鹉热嗜衣原体的实时定量PCR检测方法具有很高的特异性和敏感性,可以用于鹦鹉热嗜衣原体的检测。

A pair of specific primers targtting the major out membrane protein （momp） gene was designed and the realtime quantitative polymerase chain reaction （qPCR） was developed using SYBR Green 1for detection of Chlamydophila psittaci. Standard curve was established and the correlation coefficient was 0. 989. This method showed a high specificity and the sensitivity of this method was about 100 times higher than that of the usual PCR. It is very useful for detection of C. psittaci.