Abstract: In order to accurately quantify the content of mutton in meat products, this study established the functional relationship between copy number and mutton weight by using micro-droplet digital PCR (ddPCR). And a quantitative detection method of mutton content based on real-time fluorescence quantitative PCR (rPCR) were also developed. The result of specific experiment showed that the DNA of sheep and goat could be specifically amplified and the detection limit was 0.01 ng/μL. For the mixed samples and the commercial samples, this method could precisely quantify more than 5% of mutton contents in meat products. This method has a potential application in the detection of mutton content in meat products.
金鹭,陈传君,林华,等. 基于实时荧光PCR对肉制品中羊肉的精确定量[J]. 食品与发酵工业, 2020, 46(8): 246-253.
JIN Lu,CHEN Chuanjun,LIN Hua,et al. Accurate quantification of mutton in meat products by real-time quantitative PCR[J]. Food and Fermentation Industries, 2020, 46(8): 246-253.
O’MAHONY P J. Finding horse meat in beef products-a global problem[J]. QJM: Monthly Journal of the Association of Physicians, 2013, 106(6): 595-597.
TIAN X, WANG J, SHEN R, et al. Discrimination of pork/chicken adulteration in minced mutton by electronic taste system[J]. International Journal of Food Science & Technology, 2019, 54(3):670-678.
WASINSKI B, OSEK J. New methods of meat species identification and detection of meat adulterations[J]. Medycyna Weterynaryjna, 2013, 69(6): 348-352.
CHENG J H, CHOU H T, Lee M S, et al. Development of qualitative and quantitative PCR analysis for meat adulteration from RNA samples[J]. Food Chemistry, 2016, 192: 336-342.
EBBEHOJ E F, THOMSEN P D. Differentiation of closely related species by DNA hybridization[J]. Meat Science, 1991, 30(4): 359-366.
TARTAGLIA M, SAULLE E, PESTALOZZA S, et al. Detection of bovine mitochondrial DNA in ruminant feeds: a molecular approach to test for the presence of bovine-derived materials[J]. Journal of Food Protection, 1998, 61(5): 513-518.
AMARAL J S, SANTOS G, OLIVEIRA M B P P, et al. Quantitative detection of pork meat by EvaGreen real-time PCR to assess the authenticity of processed meat products[J]. Food Control, 2017, 72: 53-61.
KÖPPEL R, GANESHAN A, WEBER S, et al. Duplex digital PCR for the determination of meat proportions of sausages containing meat from chicken, turkey, horse, cow, pig and sheep[J]. European Food Research and Technology, 2019, 245: 853-862.
NOH E S, PARK Y J, KIM E M, et al. Quantitative analysis of Alaska pollock in seafood products by droplet digital PCR[J]. Food Chemistry,2019,275:638-643.
KÖPPEL, R, GANESHAN A, WEBER S, et al. Duplex digital PCR for the determination of meat proportions of sausages containing meat from chicken, turkey, horse, cow, pig and sheep[J]. European Food Research and Technology, 2019, 245: 853-862.
FLOREN C, WIEDEMANN I, BRENIG B, et al. Species identification and quantification in meat and meat products using droplet digital PCR (ddPCR)[J]. Food Chemistry, 2015, 173: 1 054-1 058.
SMITH C J, NEDWELL D B, DONG L F, et al. Evaluation of quantitative polymerase chain reaction-based approaches for determining gene copy and gene transcript numbers in environmental samples[J]. Environmental Microbiology, 2006, 8(5):804-815.
Codex Committee on Methods of Analysis and Sampling. Guidelines on performance criteria and validation of methods for detection identification and quantification of specific DNA sequences and specific proteins in foods: CAC/GL740-2010[S]. Rome: Codex Alimentariusliment, 2010.
THALMANN O,HEBLER J,POINAR H N,et al.Unreliable mt DNA data due to nuclear insertions: A cautionary tale from analysis of humans and other great apes[J]. Molecular Ecology, 2004, 13(2): 321-335.
MURUGAIAH C, NOOR Z M, MASTAKIM M, et al. Meat species identification and Halal authentication analysis using mitochondrial DNA[J]. Meat Science, 2009, 83(1): 57-61.
FLOREN C, WIEDEMANN I, BRENIG B, et al. Species identification and quantification in meat and meat products using droplet digital PCR (dd PCR)[J]. Food Chemistry, 2015, 173(173): 1 054-1 058.
RENÉ K, JÜRG R, JÜRG R. Multiplex real-time PCR for the detection and quantification of DNA from beef, pork, horse and sheep[J]. European Food Research and Technology, 2011, 232(1): 151-155.
HIRD H, CHISHOLM J, SANCHEZ A, et al. Effect of heat and pressure processing on DNA fragmentation and implications for the detection of meat using a real-time polymerase chain reaction[J]. Food Additives and Contaminants, 2006, 23(7): 645-650.
EBBEHOJ K F, THOMSEN P D. Species differentiation of heated meat products by DNA hybridization[J]. Meat Science, 1991, 30(3):221-234.
MARTíN I, GARCÍA T, VIOLETA F, et al. SYBR-Green real-time PCR approach for the detection and quantification of pig DNA in feedstuffs[J]. Meat Science, 2009, 82(2): 252-259.
CAI Y C, HE Y P, LV R, et al. Detection and quantification of beef and pork materials in meat products by duplex droplet digital PCR[J]. Plos one, 2017, 12(8): e091 949.
IZADPANAH M, MOHEBALI N, GORJI Z E, et al. Simple and fast multiplex PCR method for detection of species origin in meat products[J]. Journal of Food Science and Technology, 2018, 55(2): 698-703.