In order to accurately quantify the proportion of mutton-derived ingredients in meat products, the single-copy gene of mutton for quantitatively detection of mutton ingredients by the droplet digital polymerase chain reaction (ddPCR) was established in meat products. The result showed that an internal standard for quantitative analysis of the proportion of mutton-derived ingredients with ddPCR was established based on the linear relationship between DNA copy number and sample quality by adding beef as internal standard. The method developed here realized one-step transformation from target gene copy number to sample quality. The detection limit reached 0.01% of sheep-source ingredients which equaled to 0.12 copies/μL. In reality, it could accurately quantify the mutton-derived ingredients which was more than 5% of mutton in meat products. The quantitative analysis of commercial samples showed that ddPCR could accurately determine the proportion of mutton-derived ingredients in meat samples. So ddPCR had great potential in the detection and adulteration identification of mutton-derived ingredients in meat and meat products.
CHEN Chuanjun
,
JIN Lu
,
LIN Hua
,
HU Bin
,
HAN Guoquan
,
CHEN Shijie
,
ZHANG Jing
,
AN Wei
,
YANG Miao
. Quantification of mutton-derived ingredients in food by droplet digital PCR[J]. Food and Fermentation Industries, 2020
, 46(6)
: 229
-237
.
DOI: 10.13995/j.cnki.11-1802/ts.022599
[1] 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.
[2] KUMAR A, KUMAR R R, SHARMA B D, et al. Identification of species origin of meat and meat products on the DNA basis: a review[J]. Critcal Reviews in Food Science and Nutrition, 2015, 55(10): 1 340-1 351.
[3] 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.
[4] CAI Y C, HE Y Ping, LYU 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.
[5] 苗丽,张秀平,陈静,等.肉制品中羊源性成分微滴数字PCR法定量检测方法的研究[J]. 食品工业科技, 2016, 37(4): 73-76.
[6] DRUML B, MAYER W, CICHNA-MARKL M, et al. Development and validation of a Taq Man real-time PCR assay for the identification and quantification of roe deer (Capreolus) in food to detect food adulteration[J]. Food Chemistry, 2015, 178: 319-326.
[7] REN J A, DENG T T, HUANG W S, et al. A digital PCR method for identifying and quantifying adulteration of meat species in raw and processed food[J]. PLos One, 2017, 12(3): e0 173 567.
[8] CAI Y C, LI X, LYU R, et al. Quantitative analysis of pork and chicken products by droplet digital PCR[J]. Biomed Research International, 2014, 2014: 1-6.
[9] 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.
[10] 陈传君,金鹭,林华,等.干制羊肉基因组DNA不同提取方法的比较研究[J]. 核农学报,已录用,待刊.
[11] BURHANEITIN YALSINKAYA,YUMBUL E,ERKAN MOZZOGˇLU, et al. Comparison of DNA extraction methods for meat analysis[J]. Food Chemistry, 2016, 221: 1 253-1 257.
[12] 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(4): 853-862.
[13] MORISSET D, STEBIH D, MILAVEC M, et al. Quantitative analysis of food and feed samples with droplet digital PCR[J]. Plos one, 2013, 8(5): e62 583.
[14] SHEHATA H R, LI J P, CHEN S, et al. Droplet digital polymerase chain reaction (ddPCR) assays integrated with an internal control for quantification of bovine, porcine, chicken and turkey species in food and feed[J]. Plos One, 2013, 12(8): e0 182 872.
[15] 陈晨,张岩,李永波,等.微滴式数字PCR对肉制品中羊肉和猪肉定量分析[J]. 现代食品科技, 2018, 34(1): 221-227.
[16] BALLIN N Z, VOGENSEN F K, KARLSSON A H. Species determination-Can we detect and quantify meat adulteration?[J]. Meat Science, 2009, 83(2): 165-174.
[17] 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.
[18] KOPPEL R, RUF J, RENTSCH J. 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.
