title: Identification of pork in meat products using real-time loop-mediated isothermal amplification.
abstract: 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 ofcytochrome b(cytb) gene was amplified at 63 °C using SYBR Green I for 45 min 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 °C for 15 min, 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.
authors.affiliation: Changsha Center of Supervision & Inspection on Food Quality Safety , Yinshuang Road, Yuelu District, Hunan, Changsha 410013 , People's Republic of China.
authors.forename: Lixia
authors.initials: L
authors.lastname: Yang
authors.affiliation: Changsha Center of Supervision & Inspection on Food Quality Safety , Yinshuang Road, Yuelu District, Hunan, Changsha 410013 , People's Republic of China.
authors.forename: Shujun
authors.initials: S
authors.lastname: Fu
authors.affiliation: Changsha Center of Supervision & Inspection on Food Quality Safety , Yinshuang Road, Yuelu District, Hunan, Changsha 410013 , People's Republic of China.
authors.forename: Xinkai
authors.initials: X
authors.lastname: Peng
authors.affiliation: Changsha Center of Supervision & Inspection on Food Quality Safety , Yinshuang Road, Yuelu District, Hunan, Changsha 410013 , People's Republic of China.
authors.forename: Le
authors.initials: L
authors.lastname: Li
authors.affiliation: Changsha Center of Supervision & Inspection on Food Quality Safety , Yinshuang Road, Yuelu District, Hunan, Changsha 410013 , People's Republic of China.
authors.forename: Taoping
authors.initials: T
authors.lastname: Song
authors.affiliation: Changsha Center of Supervision & Inspection on Food Quality Safety , Yinshuang Road, Yuelu District, Hunan, Changsha 410013 , People's Republic of China.
authors.forename: Lin
authors.initials: L
authors.lastname: Li
keywords: cytochrome b; meat species identification; mtDNA; pork; real-time loop-mediated isothermal amplification