鸭源A型鹦鹉热衣原体的分离鉴定

Isolation and identification of genotype A Chlamydia psittaci strain from duck

目的对疑似鹦鹉热衣原体感染鸭进行检测、病原分离培养和基因型鉴定。方法从山东某鸭场采集病鸭肺脏,进行衣原体荧光PCR检测;将阳性样本匀浆和无菌处理后,接种L929细胞,连续传代培养,通过Giemsa染色和电子显微镜观察细胞内衣原体包涵体的形成;提取分离株DNA并测序,分析16S rRNA、16-23S IGS和ompA基因序列进行种属鉴定和基因分型。结果成功分离到1株鸭源鹦鹉热衣原体,基因型鉴定为A型。结论鸭源A型鹦鹉热衣原体的分离鉴定扩大了对A型菌株感染宿主范围的认识,为其生物学特性、致病性和防控技术及公共卫生等相关研究提供基础资料。

Chlamydia psittaci is a worldwide distributed zoonotic pathogen that infects a variety of birds.In order to characterization of the duck originated C.psittaci strains,lung samples were collected from suspected infection ducks and was detected by real-time PCR.The positive samples were homogenized in phosphate buffered saline with kanamycin and streptomycin,and then inoculated onto L929 cells monolayer.After several sets of passages,chlamydial inclusions of the isolate in cultured cells were observed after Giemsa staining or by electron microscope.For species identification,16S rRNA,16-23S IGS gene fragments were sequenced and analyzed.Genotyping of the isolate was performed by comparative analysis of the obtained ompA gene sequence with that of different genotype of C.psittaci strains.A C.psittaci strain was successfully isolated from the lung sample of duck by cell culture and was identified as genotype A.This study expanded our understanding of the host range of genotype A C.psittaci strain,and provided basis for further research on the pathogenicity,transmission,and public health risk of this pathogen.