鹦鹉热嗜衣原体PCR检测体系的建立

Development of Polymerase Chain Reaction to Detect Chlamydophila psittaci

鹦鹉热是由鹦鹉热嗜衣原体(Chlamydophila psittaci C.psittaci)引起,并可由带病鸟类传染给人类的一种人畜共患传染病。虽然有巢式PCR和多重PCR被开发用于C.psittaci的检测,但是这些方法较为繁琐。本研究应用了32株衣原体和嗜衣原体菌株,24株来自鸟类及人的肠道与呼吸道的细菌菌株,此外还使用606个禽鸟粪便、泄殖腔棉拭子及人体气管分泌物的临床样本。笔者以C.psittaci的MOMP基因(major outer membrane protein gene)为靶基因而开发了PCR检测方法。该最低能检测到100 fg DNA(相当于10个衣原体)相当的鹦鹉热嗜衣原体。该靶基因区域涵盖了除C.pneumoniae、C.pecorum、C.trachomatis、C.suis和C.muridarum以外的鹦鹉热嗜衣原体所有的菌株及其他部分嗜衣原体菌株。606例临床样本中有29例鸟类粪便样本、25例泄殖腔棉拭子样本和2例人源样本检测为阳性。结果表明,该PCR检测法是检测鸟源鹦鹉热嗜衣原体的快速、敏感、有效的方法。

Psittacosis is caused by Chlamydophiila psittaci （C. psittacl） and transmitted from infected birds to humans. Although nested PCR and multiples PCR home been developed for deteeting C. psittaci gene. However, these methods were not so simple and convenient. Object： The PCR besed Reaction on amplification of the major outer membrane protein （MOMP） gene of C. psittaci was developed. Methods： Thirty-two Chlamydophila and Chlamydia strains were ased in this study. Moreover, twenty- four strains of other bacteria commonly encountered in avian or human intestinal and respiratory tracts,six hundred and six avian and human clinic specimens were also used. Results： The sensitivity of the PCR was 100fg of Chlamydophilo DNA, equivalent to 10 genome copies. The target DNA region was conserved in all the Chlamydophila psittaci strains and partially conserved in other Chlomydophila species except Chlamydophila pneumoniae, Chlamydophila pecorum, Chlomydia trachomatis, Chlamydia suis and Chlamydia muridarum. Twentynine of 579 avian fecal specimens, 25 avian cloacal swabs and 2 human samples were identified as positive for C. psittaci by this PCR system. Conclusion： This newly developed PCR is a rapid, sensitive and reliable method for the detection of C. psittaci of avian origin.