Identification of pork in meat products using real-time loop-mediated isothermal amplification

被引:48
作者
Yang, Lixia [1 ]
Fu, Shujun [1 ]
Peng, Xinkai [1 ]
Li, Le [1 ]
Song, Taoping [1 ]
Li, Lin [1 ]
机构
[1] Changsha Ctr Supervis & Inspect Food Qual Safety, Changsha 410013, Hunan, Peoples R China
关键词
meat species identification; real-time loop-mediated isothermal amplification; mtDNA; cytochrome b; pork; RAPID DETECTION; PCR; CHICKEN; VIRUS; QUANTIFICATION; INFECTION;
D O I
10.1080/13102818.2014.963789
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
In this study, a one-step, real-time, loop-mediated isothermal amplification (RealAmp) assay was developed, for the highly specific detection of pork DNA. For the assay, the mtDNA of cytochrome b (cytb) gene was amplified at 63 degrees C using SYBR Green I for 45min with a Real-Time Polymerase Chain Reaction (PCR) System that measured the fluorescent signal at one-minute intervals. As little as 1 pg of template DNA could be detected, without any cross-reactivity with non-target species. Meat mixtures, heat-treated at 100 degrees C for 15min, prepared by mixing pork meat with beef at different ratios (0.01%-10%) were tested, and the RealAmp assays allowed the detection of as little as 0.01% pork in the meat mixtures. Thus, this work showed that RealAmp could be used for specific identification and sensitive quantification of meat species, even for heat-treated meat products.
引用
收藏
页码:882 / 888
页数:7
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