PCR快速检测甲型副伤寒沙门菌鞭毛抗原H-a研究

A Study on the Rapid Detection of Flagellum Antigen H-a of S. Paratyphi A by PCR

目的 研究快速、特异、灵敏的检测甲型副伤寒沙门菌鞭毛特异相H a的PCR法。方法 选择甲型副伤寒沙门菌鞭毛抗原H a基因的特异引物进行PCR扩增 ,检测甲型副伤寒沙门菌标准菌株和我省不同地区上送的 5 7株甲型副伤寒沙门菌以及非甲型副伤寒沙门菌的其它菌株 ;将甲型副伤寒沙门菌标准菌株按10 - 1 ～ 10 - 9稀释后扩增比较PCR的检测灵敏度。结果 所有甲型副伤寒沙门菌均在 3 2 9bp处出现甲型副伤寒沙门菌鞭毛抗原基因产物 ,非甲型副伤寒沙门菌株PCR结果均为阴性 ;PCR检测下限为 10 3cfu/ml。结论 PCR比传统细菌检测方法更特异、快速、灵敏、简便 ,为临床诊断、实验室筛查以及流行病学快速查源、溯源提供了新的手段。

Objective To establish a PCR assay for detection of Flagellum Antigen H a of S. paratyphi A.Methods The specific primers of flagellum antigen H a of S. paratyphi A were selected to be amplified by PCR assay to detect the S. paratyphi A of standard kind and 57 kinds of S. paratyphi A and other kind of non S. paratyphi A from different area in Zhejiang province. The sensitivity of PCR assay was analyzed with 10 -1 ～10 -9 diluted. Results The gene product of flagellum antigen H a was detected in all S. paratyphi A at 329bp, but was not detected in non S. paratyphi A. The lower limit of PCR detection was 103cfu/ml. Conclusions PCR assay is better than traditional detection for its specificity, celerity,sensitivity and convenience and it is helpful in clinical diagnosis, laboratory screen and epidemiological investigation.