基于多重PCR的主要食源性致病菌的快速检测  

Rapid detection of foodborne pathogenic bacteria based on multiple PCR

作　　者：胡瑞丽[1] 任方方[1] 吴洋洋[1] 许燕[1] 赵凯[2,3] 索玉娟[4,5] 邵毅[4,5] 周昌艳[4,5] 

机构地区：[1]上海师范大学,上海200234 [2]上海市农业科学院生物技术研究所,上海201106 [3]上海市农业遗传育种重点实验室,上海201106 [4]上海市农业科学院农产品质量标准与检测技术研究所,上海201403 [5]上海科立特农产品检测技术服务有限公司,上海201403

出　　处：《上海农业学报》2015年第4期34-39,共6页Acta Agriculturae Shanghai

基　　金：上海市闵行区产学研项目(2013MH099);上海市科技新农重点攻关项目[沪农科攻字(2013)第3-2号]

摘　　要：根据单核增生李斯特菌、沙门氏菌、金黄色葡萄球菌、大肠杆菌0157：H7的特异性序列分别设计4对特异性引物,建立了一种快速检测上述4种常见食源性致病菌的多重PCR方法,并对反应体系中的模板、引物、rTaq酶、MgSO_4添加量以及退火温度进行了优化。结果显示：4对引物序列的特异性均较好,利用单重PCR对4种致病菌的灵敏度进行检测,其检测限均达到10~3copies/mL。多重PCR同时检测4种致病菌的灵敏度为10~3copies/ml,对人工污染猪肉中的4种致病菌的灵敏度检测为10~3cfu/mL。该检测方法不与其他菌产生交叉反应,与传统方法相比大大缩短了检测时间,并提高了检测的准确度。According to the specific sequence of 4 common foodborne pathogenic bacteria, Listeria monocy- togenes, Salmonella enterica, Staphylococcus aureus and Escherichia coli O157 ：H7,4 pairs of specific primers were designed and a multiple PCR method for rapid detecting the above bacteria was established. The additive amount of template, primers, rTaq, MgSOg, and the annealing temperature of the reaction system were optimized. The re- suits showed that the specificity of 4 pairs of primers was better. The sensitivity of 4 species of pathogenic bacteria was detected by single PCR, and the detection limit was 103copies/mL. The sensitivity of multiple PCR for simul- taneous detection of 4 species of pathogenic bacteria was 103copies/mL, and the sensitivity of 4 species of patho- genic bacteria in artificial contaminated pork was detected as 103cfu/mL. The detection method did not produce cross reactions with other bacteria. Compared with the traditional method, the detection time was greatly short- ened, and the accuracy of detection was improved.

关 键 词：多重PCR 食源性致病菌 检测 食品安全 

分 类 号：S853.31[农业科学—临床兽医学]