空肠和结肠弯曲菌鞭毛蛋白基因fla A的检测

Detection of fla A of Campylobacter jejuni and Campylobacter coli by PCR

目的运用一种快速、敏感、特异的检测空肠和结肠弯曲菌的方法。方法以空肠和结肠弯曲菌所共有特异的鞭毛蛋白基因flaA的一段高度保守序列为引物,用PCR法扩增flaA基因上的一段约1700bp的片断。用该引物对空肠和结肠弯曲菌的标准株、福建省的食品分离株进行PCR扩增检测,并同时检测该PCR方法的敏感性。结果扩增片断表现出极好的特异性,2株空肠和结肠弯曲菌标准菌株、8株分离自不同食品样品的空肠弯曲菌和结肠弯曲菌菌株均为阳性,且敏感性实验显示该PCR方法的反应体系最低检出菌量为6CFU。结论该方法快速、敏感、特异,可用于突发性食物中毒和暴发感染的调查。

Objective To seek a fast, sensitive and specific method for detection of C. jejuni and C. coli in foods. Method The highly conserved consensus sequences in flaA of C. jejuni and C. coli were selected as primers to amplify a 1 70Obp fragment in the standard strains of C. jejuni and C. coli and the isolated strains from foods in Fujian Province. Results The sensitivity of PCR was determined. Specific fragments were obtained from standard strains of C. jejuni and C. coli, 4 isolated srtains of C. jejuni and 4 isolated strains of C. coli from foods. A minimum of 6 CFU could be detected in the PCR reaction system. Conclusion Detection of flaA by PCR could detect C. jejuni and C. coli rapidly with high sensitivity and specificity and could be applied for investigations during emergency of food poisoning and outbreak of infection.