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乳制品中水牛成分的实时荧光PCR检测方法
李富威1,张舒亚2,曾庆坤1,王赢1,袁辰刚1,王珏1,林波1,包建强1
1.
2. 上海出入境检验检疫局
Real-time PCR for the detection buffalo ingredients in dairy products
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摘要 水牛奶制品因其良好的营养功效备受国内外消费者的青睐,而在国内外,乳制品掺假行为,却常有发生。我国作为水牛奶酪等乳制品的进口国,迫切需要一种准确的水牛成分的检测方法,来避免这种掺假行为对我国的进出口贸易造成损失。根据水牛(Bubalus bubalus)线粒体细胞色素b(Cytb)基因的保守序列,设计一对特异性引物和TaqMan-MGB探针,建立了水牛乳成分的实时荧光PCR检测方法。运用实时荧光PCR方法对5种乳水牛样品和47种常见的非水牛动植物样品进行检测,5种水牛乳样品均产生荧光信号,其余非水牛样品均不产生荧光信号,表明该检测方法具有特异性。该检测方法的重量检测灵敏度为0.01%水牛奶(W/W),DNA浓度检测灵敏度为0.001 ng/μL水牛DNA。对市售的水牛奶制品进行实际样品检测,结果表明,该方法能很好地检出水牛乳成分。本研究表明,该方法具有特异性好、灵敏度高的优点,可作为乳制品中水牛乳成分鉴别检测的有效方法。
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李富威
张舒亚
曾庆坤
王赢
袁辰刚
王珏
林波
包建强
关键词 水牛实时荧光PCRTapMan-MGB探针Cytb基因检测    
Abstract:Dairy product authenticity is presently a subject of great concern to food authorities, as the incorrect labelling of foodstuffs can present a commercial fraud. Many consumers are concerned about dairy products they eat and accurate labelling is important to inform consumer choice. Thus, the detection of milk species is important in cheese making, not only with regards to accurate consumer information and legal aspects (concerned with labeling and guarantee requirements) but also to medical requirements, food allergies or religious practices. Specific primers and Taq Man-MGB probe were designed according to highly conserved sequence of mitochondrial cytochrome b(Cytb) gene of buffalo(Bubalus bubalus) to develop a Real-time fluorescence polymerase chain reaction (Real-time PCR) assay for rapid detection of buffalo ingrediens. The primers and probes could specific identify 5 kinds of buffalo with 47 kinds of samples. The detection limit of the method was 0.001 ng/μL DNA concentration and 0.01 %(W/W) buffalo milk. The commercial samples were detected by Real-time PCR and the detection results showed that buffalo ingredients were detected well. All the results confirm that the Real-time PCR assay is a sensitive and specific method for the detection of buffalo materials in dairy products.
Key wordsBuffalo    Real-time PCR    TaqMan-MGB probe    Cytb gene    Detection
收稿日期: 2012-06-13     
通讯作者: 张舒亚   
引用本文:   
李富威1,张舒亚2,曾庆坤1,王赢1,袁辰刚1,王珏1,林波1,包建强1. 乳制品中水牛成分的实时荧光PCR检测方法[J]. , 2013, 21(2): 247-252.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2013/V21/I2/247
 
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