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食品中4种致病微生物的多重PCR快速检测技术研究
引用本文:杨平,杨迎伍,陈伟,王国民,李正国.食品中4种致病微生物的多重PCR快速检测技术研究[J].西南农业大学学报,2007,29(5):90-94.
作者姓名:杨平  杨迎伍  陈伟  王国民  李正国
作者单位:重庆大学基因工程研究中心、重庆大学生物工程学院、重庆市高校功能基因与调控新技术重点实验室,重庆400044
基金项目:中法先进研究计划(PRABT-04-01),重庆市科委自然基金资助项目(No.8479).
摘    要:传统微生物检测方法耗时长、灵敏性差,难以满足食品安全快速检测要求,建立和完善食品中致病微生物快速检测技术具有重要的现实意义。本研究针对沙门氏菌、志贺氏菌、金黄色葡萄球菌和单核细胞增生性李斯特菌,根据其特异基因设计出4对引物,通过对食品样品进行梯度离心处理、改良异硫氰酸胍法提取基因组DNA和多重PCR(multiplex PCR)条件的优化,建立了这4种食源性致病微生物的多重PCR检测方法。通过人工污染检测和市场样品检测,同时进行传统方法验证,结果表明:该检测方法简单、快速、准确、灵敏度高,只需经过增茵4~8h其最低检出限可达1cfu/mL,具有较强的实际应用价值,可广泛应用于食品卫生检测和临床检验等领域。

关 键 词:食源性致病微生物  多重PCR  快速检测
文章编号:1000-2642(2007)05-0090-05
修稿时间:2007-03-28

Study on the Rapid Detection of Four Food-Borne Pathogens in Food by Multiplex PCR
YANG Ping, YANG Ying-wu, CHEN Wei, WANG Guo-ming, LI Zheng-guo.Study on the Rapid Detection of Four Food-Borne Pathogens in Food by Multiplex PCR[J].Journal of Southwest Agricultural University,2007,29(5):90-94.
Authors:YANG Ping  YANG Ying-wu  CHEN Wei  WANG Guo-ming  LI Zheng-guo
Institution:Genetic Engineering Research Center, Bio-Engineering College, Chongqing University, Key Lab of Functional Gene and New Regulation Technologies under Chongqing Municipal Education Commission, Chongqing 400044, China
Abstract:The traditional detection methods of food-borne pathogens are often time-consuming and not sensitive enough. It is, hence, necessary to develop rapid technologies for the detection of food-borne pathogenic microorganisms. According to the sequence of special pathogens' target genes of four food-borne pathogens: Salmonella, Shigella, Staphylococcus aureus and Listeria monocytogenes, four pairs of specific oligonucleotide primers were designed. Food samples was pre-treated by gradient centrifugation, and genome DNA was extracted by the guanidine thiocyanate. After optimizing the reaction system of multiplex PCR, a rapid simultaneous detection method for the four food-borne pathogens was established. Artificially contaminated food and marketable samples were inspected with this method. The results showed that this method was simple, rapid, sensitive and applicative, and the detection limit was as low as 1 cfu/mL after 4 to 8 hours' culture. This multiplex PCR method could be used in the fields such as food sanitation detection and clinical inspection.
Keywords:food-borne pathogenic microorganism  multiplex PCR  rapid detection
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