摘要
以16S rRNA、mapA和ceuE为靶基因,建立检测空肠弯曲菌和结肠弯曲菌的多重PCR方法。特异性试验能分别扩增出空肠弯曲菌和结肠弯曲菌的特异性条带,其他参考菌株均未扩增出条带;敏感性试验表明,基于增菌培养和选择性培养的多重PCR方法对弯曲菌检测限为10 mL或10 g模拟污染样品,最低可检测0.925 pg.μL-1空肠弯曲菌DNA和1.050 pg.μL-1结肠弯曲菌DNA。以国家标准为参照,应用该方法对180份生鲜鸡肉样品进行检测,结果表明多重PCR方法具有快速、简便、特异性强和灵敏度高等特点,可克服传统生化鉴定和血清学方法的不足,为弯曲菌检测提供新的技术。
A multiplex PCR （m-PCR） targeting 16,5 rRNA gene, mapA gene and ceuE gene for the detection of Campylobacter jejuni and Campylobacter coli was developed. Species-specific product could be detected after amplification of the DNA template of C. jejuni and C. coli, while other bacteria strains could not be detected. The m-PCR assay could detect as low as 10 mL or 10 g, artificial contaminated samples and its detect limit of C. jejuni and C. coli were 0. 925 and 1. 050 pg · uL^-1 DNA template respectively. 180 of fresh chicken .samples were detected for the C. jejuni and C. coli by the m- PCR, and the result was coincided with that of the national standard method for Campylobacter. It was suggested that the m-PCR was highly specific and sensitive, and could overcome the shortcomings of biochemical and serological identification.
出处
《扬州大学学报（农业与生命科学版）》
CAS
CSCD
北大核心
2007年第1期5-8,共4页
Journal of Yangzhou University：Agricultural and Life Science Edition
基金
国家杰出青年科学基金资助项目(30425031)
中丹合作项目(AM14:22NNP34)
教育部FANEDD专项资金资助项目(200358)
关键词
空肠弯曲菌
结肠弯曲菌
多重PCR
快速检测
Campylobacter jejuni
Campylobacter coli
multiplex PCR
rapid detection
