三种方法对禽鹦鹉热嗜性衣原体疑似病例病原的流行调查

目的利用免疫荧光抗体法、PCE-ELISA、PCR法分别检测北京市及周边地区禽鹦鹉热嗜性衣原体疑似病例的病原,以了解和评价该病原流行状况。方法收集临床疑似禽衣原体的父母代、商品代家禽、SPF鸡喉头拭子、气囊组织、子宫黏膜,组织标本固定后采用直接免疫荧光染色测定衣原体包含体;喉头拭子、子宫黏膜处理后以PCE-ELISA定量测定衣原体;以衣原体保守性高的序列设计CTU/CTL引物扩增病料组织的omp-1基因片段。结果直接荧光染色法显示家禽衣原体平均阳性率为38.7%,组织检出率依次为子宫黏膜、气囊、喉头拭子;PCE-ELISA检测显示平均阳性率为58.7%,其中SPF鸡阳性率为10.0%,健康肉鸡达到30.0%;PCR法检测家禽衣原体平均阳性率为71.6%,SPF鸡为10.0%。样本的目的基因与标准禽衣原体6BC同源性超过99%。结论种禽场和商品养殖场均发现疑似病例中禽鹦鹉热嗜性衣原体感染情况较为严重。荧光抗体染色、PCE-ELISA可用于临床实践中进行快速、准确检测。

Prevalence of Avian Chlamydophila psittaci in Suspected Cases Using Three Tests

Objective To detect the pathogen of avian Chlamydophila psittaci in suspected poultry cases in order to investigate the prevalence of chlamydophilosis. Methods Throat swabs, air sacs and uterine mucosa from suspected parent birds, commercial hens, broilers, ducks and specific pathogen free （SPF） hens were collected and sampled appropriately. The fixed samples were examined to detect the specific inclusion bodies by a direct immunofluorescence test. Meanwhile, the clinical samples were subjected to detect the antigen positivity by PCE-ELISA and PCR method targeting omp-1 gene. Results In the current study, an average positivity of 38.7% was confirmed by immunofluorescence test. An average positivity of 58.7% was detected by PCE-ELISA, including 10.0% in SPF hens and 30.0% in clinical healthy broilers. Moreover, out of 60 clinical samples, 71.6% of samples was positive for target gene, particularly, 10% of the samples was detected to be positive in SPF layers. The tested gene sequences showed more than 99.0% similarity to that of reference C. psittaci of 6BC. Conclusion Avian chlamydophilosis is prevalent in beth parent birds and in commercial flocks by the three tests. Immunofluorescence test and PCEELISA are proved to be two fast and reliable methods for investigating epidemiology of avian Chlamydophila psittaci in field.