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PCR法和培养法调查食品和水中空肠弯曲杆菌的比较研究

Research on survey of Camplobater jejuni in food and water by PCR and culture methods

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【作者】 阳成波蒋原黄克和祝长青

【Author】 YANG Chengbo,JIANG Yuan,HUANG Kehe,ZHU Changqing (College of Veterinary Medicine Nanjing Agricultural University,Nanjing,210095)

【机构】 南京农业大学动物医学院江苏出入境检验检疫局江苏出入境检验检疫局 南京210095南京210095

【摘要】 目的 PCR方法和培养法调查食品和水中空肠弯曲杆菌的比较研究。方法 用空肠弯曲杆菌VS1基因的序列为模板设计一对引物 ,建立一种快速从食品和水中检测空肠弯曲杆菌的PCR方法 ;同时对南京地区的屠宰场、超市和牛奶场等地采取 10 0份鸡肉、10 0份生牛奶及 10 0份井水及自来水进行空肠弯曲杆菌PCR法和培养法双向检查。结果 研究表明该PCR方法只对空肠弯曲杆菌能特异的扩增出 35 8bp片段 ,而其它结肠弯曲杆菌、胎儿弯曲杆菌、霍乱弧菌、创伤弧菌、沙门氏菌、大肠杆菌等均不能扩增出 ,检测灵敏度达 8CFU/ml,特异性与常规生化检查是一致的 ,发现 30 % (30 / 10 0 )的鸡肉为阳性、30 % (30 / 10 0 )的生牛奶为阳性、14 % (14 / 10 0 )的水为阳性。结论 调查结果表明在市场上销售的鸡肉、牛奶、牛奶制品及井水存在不同程度空肠弯曲杆菌污染 ,如果加工不当或饮食习惯不卫生 ,会对消费者的健康构成潜在威胁。

【Abstract】 Aim To research and survey the Camplobater jejuni in food and water by PCR and culture methods Methods A polymerase chain reaction (PCR) method designed to sensitively detect and identify C jejuni is described The species special sequence of VS1 was amplified and characterized with a pair primer As an application of the PCR system and API Campy,a set of 100 frozen chicken meat samples,100 milk samples and 100 water samples was screened for the presence of C jejuni Results A total of 20 bacterial strains,8 of C jejuni and 12 of other species,were tested,and giving no false positive or false negative results The detection limit as determined by ethidium bromide staining of amplification products on agarose gels was 8 bacteria or less in artificially contaminated water,milk,and poultry samples with the primer PCR assay 30% (30/100) of chicken meat samples,30% (30/100) of milk samples and 14% (14/100) of water samples were found to be positive for C jejuni Conclusion These results support the assertion that retail chicken meat,raw milk and environmental water is commonly contaminated with C jejuni,and pose a potential risk to consumer if hygiene and cooking are not adequate

【关键词】 聚合酶链反应空肠弯曲杆菌食品污染
【Key words】 PCRCampylobacter jejunifoodwatercontamination
  • 【文献出处】 中国人兽共患病杂志 ,Chinese Journal of Zoonoses , 编辑部邮箱 ,2003年01期
  • 【分类号】R155.5
  • 【被引频次】29
  • 【下载频次】296
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