A sensitive multiplex PCR protocol for simultaneous detection of chicken, duck, and pork in beef samples
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A rapid and sensitive multiplex PCR assay was developed for simultaneous identification of the adulteration ingredients of chicken, duck and pork in beef. Specific primers for the mitochondrial genes of Cyt b, CO III, ATPase subunit 8/6 and Cyt b of chicken, duck, pork, and beef, respectively, were adopted in the assay. DNA exaction from meat samples was carried out by using magnetic nanoparticles as rapid separation substrates. The multiplex PCR assay showed that the limit of detection was 0.05% for each species. Moreover, the multiplex PCR specifically identified five beef samples adulterated with pork and one beef samples adulterated with chicken among the 35 commercial samples examined, indicating the practicability of this multiplex PCR method for identifying adulterated ingredients of chicken, duck, and pork in commercial beef products.
KeywordsMultiplex PCR Identification Chicken Duck Pork Beef Meat adulteration Sensitivity Specificity
This work is financially supported by the Grant of 2017YFF0208600, China Agriculture Research System-48 (CARS-48), Anhui Provincial Modern Argo-industry Tech. Research System (NYCYTX-2016-84), NSFC 21475030 and the National 10000 Talents-Youth Top-notch Talent Program.
- Choi SJ, Hur GY, Shin SY, Park HS (2007) A case of adult onset cow’s milk allergy presenting beef and pork meat allergy. Korean J Asthma Allergy Clin Immunol 27(3):200–203Google Scholar
- Fan L, Li P, Ding H, Jin P, Fu C (2013) Detection for bovine-derived ingredients in foods with real-time polymerase chain reaction method. Sci Technol Food Ind 34:65–67 (in Chinese) Google Scholar
- González-Mancebo E, Pastor C, González-de-Olano D, Gandolfo-Cano M, Melendez A, Cuesta J, Zapatero A, Bartolomé B (2011) Identification of allergens in chicken meat allergy. J Investig Allergol Clin Immunol 21:326–327Google Scholar
- Li L, Xu J, Zhang Y, Wang Z (2004) Identification of components from animal speaies in meat and bone meals by PCR. Chin Anim Health Insp 21:29–31 (in Chinese) Google Scholar
- Mutalib SA, Muin NM, Abdullah A, Muin M, Abdullah A, Hassan O, Mustapha WAW, Sani NA, Maskat MY (2015) Sensitivity of polymerase chain reaction (PCR)-southern hybridization and conventional PCR analysis for Halal authentication of gelatin capsules. LWT Food Sci Technol 63:714–719CrossRefGoogle Scholar
- Zhang J, Zong H, Zhang L (2008) PCR-mtDNA for detecting components of duck origin in foodstuff and feedstuff. Chin J Biotechnol 24:1832–1836Google Scholar
- Zhang Y, Zhu Z, Yang W, Ren J, Tan X, Wang Y, Mao N, Xu S, Zhu S, Cui A, Zhang Y, Yan D, Li Q, Dong X, Zhang J, Zhao Y, Wan J, Feng Z, Sun J, Wang S, Li D, Xu W (2010) An emerging recombinant human enterovirus 71 responsible for the 2008 outbreak of hand foot and mouth disease in Fuyang city of China. Virol J 7:1–9CrossRefGoogle Scholar