实时荧光聚合酶链式反应法检测食品中猪源性成分

doi:10.7506/spkx1002-6630-201308048

食品科学 ›› 2013, Vol. 34 ›› Issue (8): 224-227.doi: 10.7506/spkx1002-6630-201308048

实时荧光聚合酶链式反应法检测食品中猪源性成分

范丽丽，李培，傅春玲，丁洪流，陈英

1.苏州大学医学部公共卫生学院，江苏苏州 215123；2.苏州市产品质量监督检验所，江苏苏州 215128

收稿日期:2012-01-09 修回日期:2013-03-17 出版日期:2013-04-25 发布日期:2013-05-07
通讯作者: 范丽丽 E-mail:letitbe2010214@163.com
基金资助:
苏州市科技支撑计划项目

Detection for Pig-Derived Components in Foods by Real-Time Polymerase Chain Reaction

FAN Li-li，LI Pei，FU Chun-ling，DING Hong-liu，CHEN Ying

1. School of Public Health, Medical College of Soochow University, Suzhou 215123, China；2. Suzhou Products Quality Supervision and Inspection Institute, Suzhou 215128, China

Received:2012-01-09 Revised:2013-03-17 Online:2013-04-25 Published:2013-05-07

摘要/Abstract

摘要： 针对猪线粒体细胞色素b基因序列设计特异的引物和探针，建立食品中猪源性成分实时荧光聚合酶链式反应检测方法，并经特异性和敏感性试验验证其可行性。结果表明：该体系可扩增猪肉DNA片段，长度为98bp，其他常见畜、禽肉成分均无法正常扩增。该体系灵敏度低至1pg，且阴性样品扩增后的Ct值限制在35循环以后。对于各模拟肉类样品中掺杂的猪源性成分，其检测限均达1%，经市售加工食品检测验证，表明所建立的猪引物探针体系具有特异性好、灵敏度高、快速、高效等优点，可用于对食品中猪源性成分的掺假鉴别检测。

关键词: 实时荧光聚合酶链式反应, 猪, 线粒体细胞色素b基因, 食品掺假

Abstract: Species-specific real-time PCR (TaqMan) assay was developed for the detection of pig-derived components in foods. Primers and Taqman probe of this assay were designed with pig conservative regions of the mitochondrial cytochrome b (Cytb) gene (amplicon = 98 base pairs). The specificity was evaluated and achieved without amplification on DNA from other meats. Results showed that this method revealed a high sensitivity and could detect 1 pg of pork template DNA, while Ct (cycle threshold) values of negative samples were limited to 35 cycles. Applying this assay method, the recovery rate of DNA extracted from meat mixtures was up to 1% pork spiked in other species. Therefore, it is a potentially reliable and suitable technique in routine food analysis for the detection of pork in foods.

Key words: real-time polymerase chain reaction, pig, mitochondrial cytochrome b gene, food adulteration

中图分类号:

TS207.3

范丽丽，李培，傅春玲，丁洪流，陈英. 实时荧光聚合酶链式反应法检测食品中猪源性成分[J]. 食品科学, 2013, 34(8): 224-227.

FAN Li-li，LI Pei，FU Chun-ling，DING Hong-liu，CHEN Ying. Detection for Pig-Derived Components in Foods by Real-Time Polymerase Chain Reaction[J]. FOOD SCIENCE, 2013, 34(8): 224-227.

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实时荧光聚合酶链式反应法检测食品中猪源性成分

Detection for Pig-Derived Components in Foods by Real-Time Polymerase Chain Reaction

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