PCR方法快速检测食品中的金黄色葡萄球菌  

Staphylococcus aureus in food determined by polymerase chain reaction

作　　者：田静[1] 计融[1] 杨军[2] 李业鹏[1] 刘秀梅[1] 

机构地区：[1]中国疾病预防控制中心营养与食品安全所,北京100021 [2]中国医科大学公共卫生学院

出　　处：《卫生研究》2007年第2期183-186,共4页Journal of Hygiene Research

基　　金：国家科技部十五攻关重大项目(No2001BA804A03)

摘　　要：目的建立聚合酶链反应(PCR)方法快速检测牛奶、冰淇淋及肉中的金黄色葡萄球菌(金葡菌)。方法针对金葡菌耐热核酸酶基因(nuc)、纤维蛋白原结合蛋白基因(ClfA)设计并合成2对引物进行PCR扩增,特异的检测金葡菌,并对52株金葡菌和31株非金葡菌进行扩增,验证方法的特异性。在牛奶、冰淇淋及肉中人工污染不同菌量,增菌培养,在不同时间段提取DNA进行PCR扩增,确定该方法在食品中的检出限。结果nuc、ClfA两对引物对金葡菌的检出均有很好的特异性,在3种食品中的检出限均为10cfu/g(ml),全部检测可在24h完成。结论建立了适用于牛奶、冰淇淋及肉中的金葡菌检测的快速、灵敏、特异的PCR方法。Objective To establish a rapid polymerase chain reaction （PCR）method for detection of staphylococcus aureus in milk, ice cream and meat. Methods Two pairs of oligonucleotide primers were designed with thermo nuclease gene nuc and surfaced-associated fibrinogen-hinding protein gene ClfA to detect staphylococcus aurens. Fifty-two staphylococcus aureus and thirty-one non staphylococcus aureus were amplified by PCR to verify the specificity. Various numbers of bacteria were added into milk, ice cream and meat. After enrichment, DNA extracted in different time was amplified by PCR to verify detection limit. Results Each primer pair allows specific detection. The limit of detection was 10cfu/g（ml） in three kinds of food. Whole procedure of detection could be finished in 24 hours. Conclusion A rapid, sensitive and specific PCR method can be applied detecting staphylococcus aureus in milk, ice cream and meat.

关 键 词：聚合酶链反应 金黄色葡萄球菌 食品 微生物 

分 类 号：R155.51[医药卫生—营养与食品卫生学] TS207.4[医药卫生—公共卫生与预防医学]