GNM C7-8实时荧光PCR系统对掺假肉制品中多种动物源性成分的同时检测
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摘要
应用GNM C7-8实时荧光PCR系统,同时快速检测掺假肉制品中多种动物源性成分。将牛肉粉、驴肉粉、鸵鸟肉粉和羊肉粉4组不同肉粉中分别混入猪肉粉、鸭肉粉、马肉粉、鸡肉粉和狐狸肉粉中的2～3种肉粉,制备人工模拟掺假肉制品。通过GNM C7-8实时荧光PCR和ABI7500荧光PCR方法,对上述四种掺假肉制品进行多种动物源性成分检测和比较。结果表明,GNM C7-8实时荧光PCR方法与ABI7500荧光PCR方法从掺假肉制品中检测猪、鸭、马、鸡和狐狸5种成分的循环数一致,分别是31、31、29、32和25,但基于八模块设计的GNM C7-8实时荧光PCR方法能同时检测四种掺假肉制品中不同动物源性成分,检测时间更短。因此,GNM C7-8实时荧光PCR系统的八个模块能够独立运行,互不干扰,无交叉污染,能够实现对掺假肉制品中多种动物源性成分的同时快速检测,为肉制品掺假检测提供强有力的方法支持。
The GNM C7-8 real-time fluorescent PCR system was used to simultaneously and rapidly detect various animalderived ingredients in adulterated meat products.Bovine meal powder,donkey meal powder,ostrich meal powder and ovine meal powder were respectively mixed with two or three kinds of other animal meal powders of the porcine,duck,horse and chicken.Four groups of artificial adulterated meat products were prepared. They were detected and compared with various animal-derived ingredients by GNM C7-8 real-time fluorescent PCR and ABI 7500 fluorescent PCR. The results showed that the number of cycles detected by GNM C7-8 real-time fluorescence PCR for five kinds of ingredients of pigs,ducks,horses,chickens and foxes from the adulterated meat products was the same as that detected by ABI7500 fluorescent PCR method and was 31,31,29,32 and 25 respectively. However,GNM C7-8 real-time fluorescent PCR based on eight modules could simultaneously detect different animal-derived components in four adulterated meat products,and the detection time was shorter.Therefore,the eight modules of GNM C7-8 real-time fluorescent PCR system could run independently withoutinterference and cross-contamination and realize simultaneously and rapidly detection of various animal-derived ingredients in adulterated meat products,which would provide powerful method support for the detection of adulterated meat products.
The GNM C7-8 real-time fluorescent PCR system was used to simultaneously and rapidly detect various animalderived ingredients in adulterated meat products.Bovine meal powder,donkey meal powder,ostrich meal powder and ovine meal powder were respectively mixed with two or three kinds of other animal meal powders of the porcine,duck,horse and chicken.Four groups of artificial adulterated meat products were prepared. They were detected and compared with various animal-derived ingredients by GNM C7-8 real-time fluorescent PCR and ABI 7500 fluorescent PCR. The results showed that the number of cycles detected by GNM C7-8 real-time fluorescence PCR for five kinds of ingredients of pigs,ducks,horses,chickens and foxes from the adulterated meat products was the same as that detected by ABI7500 fluorescent PCR method and was 31,31,29,32 and 25 respectively. However,GNM C7-8 real-time fluorescent PCR based on eight modules could simultaneously detect different animal-derived components in four adulterated meat products,and the detection time was shorter.Therefore,the eight modules of GNM C7-8 real-time fluorescent PCR system could run independently withoutinterference and cross-contamination and realize simultaneously and rapidly detection of various animal-derived ingredients in adulterated meat products,which would provide powerful method support for the detection of adulterated meat products.
引文
[1]陈颖,吴亚君,徐宝梁,等.进出口动物源性产品中牛羊成分的检测方法[J].食品工业科技,2004(8):144-146.
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[10]Chen A,Wei C,Chen G,et al.Duplex PCR approach for the detection and quantification of donkey,horse and mule in raw and heat-processed meat products[J].International Journal of Food Science&Technology,2015,50(3):834-839.
[11]GB/T 25165-2010明胶中牛、羊、猪源性成分的定性检测方法:实时荧光PCR法[S].北京:中国标准出版社,2010.
[12]SN/T 3730 3-2013食品及饲料中常见畜类品种的鉴定方法第3部分:狐狸成分检测-实时荧光PCR法[S].北京:中国标准出版社,2013.
[13]SN/T 3730 4-2013食品及饲料中常见畜类品种的鉴定方法第4部分:驴成分检测-实时荧光PCR法[S].北京:中国标准出版社,2013.
[14]SN/T 3730 5-2013食品及饲料中常见畜类品种的鉴定方法第5部分:马成分检测实时荧光PCR法[S].北京:中国标准出版社,2013.
[15]金萍,结莉,陆俊,等.Taqman探针荧光聚合酶链式反应实时同步鉴定动物源性食品中猪肉、鸡肉源性成分[J].肉类研究,2016,30(9):17-22.
[16]侯东军,韩合敬,郝智慧,等.三重荧光PCR法鉴定食品中牛、猪、鸭3种动物源性成分[J].畜牧与兽医,2016,48(4):45-48.
[17]曾少灵,秦智锋,阮周曦,等.多重实时荧光PCR检测牛、山羊和绵羊源性成分[J].生物工程学报,2009,25(1):139-146.
[18]黄银花,胡晓湘,徐慰倬,等.影响多重PCR扩增效果的因素[J].遗传,2003,25(1):65-68.
[19]王金凤,王建昌,张伟,等.GNM C7-8实时荧光定量PCR同时快速检测8种食源性致病菌[J].食品安全质量检测学报,2018,9(9):2090-2095.
[20]Q/DXJNM0001-2012 GNM C7-8实时荧光定量PCR分析仪[S].北京:中国标准出版社,2012.
[21]李静,段永生,王建昌,等.实时荧光单引物等温扩增检测阪崎克罗诺杆菌方法的建立[J].中国食品卫生杂志,2015,27(5):524-530.
[22]李瑞,王建昌,李静,等.实时荧光单引物等温扩增(SPIA)技术检测大肠杆菌O157的方法研究[J].现代食品科技,2016,32(2):317-322,316.
[23]朱业培,王玮,吕青骎,等.基于基因芯片技术检测6种动物源性成分[J].南京农业大学学报,2015,38(6):1003-1008.
[24]吴鑫,章志超,王栋,等.基于膜基因芯片技术的肉制品中5种动物源性成分的鉴定[J].现代预防医学,2018,45(10):1762-1765,1786.
[25]Wu Q,Xiang S,Wang W,et al.Species identification of fox-,mink-,dog-,and rabbit-derived ingredients by multiplex PCRand real-time PCR assay[J].Applied Biochemistry&Biotechnology,2017(1):1-12.
[26]Rentsch J,Weibel S,Ruf J,et al.Interlaboratory validation of two multiplex quantitative real-time PCR methods to determine species DNA of cow,sheep and goat as a measure of milk proportions in cheese[J].European Food Research&Technology,2013,236(1):217-227.
[2]吕程序,韩鲁佳,杨增玲.显微镜红外在饲料中动物源性成分检测上的应用[C].//山西:纪念中国农业工程学会成立30周年暨中国农业工程学会学术年会,2009:1-7.
[3]Hossner K L,Yemm R S,Sonnenshein S E,et al.Comparison of immunochemical(enzyme-linked immunosorbent assay)and immunohistochemical methods for the detection of central nervous system tissue in meat products[J].Journal of Food Protection,2006,69(3):644-650.
[4]Barai B K,Nayak R R,Singhal R S,et al.Approaches to the detection of meat adulteration[J].Trends in Food Science&Technology,1992,3(92):69-72.
[5]李宗梦,赵良娟,赵宏,等.肉及肉制品动物源性成分鉴别技术研究进展[J].食品研究与开发,2014,35(18):122-127.
[6]石盼盼,李旭,魏法山,等.肉类掺假的分子生物学检测[J].食品与生物技术学报,2017,36(7):773-777.
[7]Abdulmawjood A,Grabowski N,Fohler S,et al.Development of loop-mediated isothermal amplification(LAMP)assay for rapid and sensitive identification of ostrich meat[J].PloS One,2014,9(6):e100717.
[8]Shi Y,Feng Y,Xu C,et al.Loop-mediated isothermal amplification assays for the rapid identification of duck-derived ingredients in adulterated meat[J].Food Analytical Methods,2017,10(7):1-7.
[9]6)Ilhak O 6)I,Arslan A.Identification of meat species by polymerase chain reaction(PCR)technique[J].Turkish Journal of Veterinary&Animal Sciences,2007,31(3):159-163.
[10]Chen A,Wei C,Chen G,et al.Duplex PCR approach for the detection and quantification of donkey,horse and mule in raw and heat-processed meat products[J].International Journal of Food Science&Technology,2015,50(3):834-839.
[11]GB/T 25165-2010明胶中牛、羊、猪源性成分的定性检测方法:实时荧光PCR法[S].北京:中国标准出版社,2010.
[12]SN/T 3730 3-2013食品及饲料中常见畜类品种的鉴定方法第3部分:狐狸成分检测-实时荧光PCR法[S].北京:中国标准出版社,2013.
[13]SN/T 3730 4-2013食品及饲料中常见畜类品种的鉴定方法第4部分:驴成分检测-实时荧光PCR法[S].北京:中国标准出版社,2013.
[14]SN/T 3730 5-2013食品及饲料中常见畜类品种的鉴定方法第5部分:马成分检测实时荧光PCR法[S].北京:中国标准出版社,2013.
[15]金萍,结莉,陆俊,等.Taqman探针荧光聚合酶链式反应实时同步鉴定动物源性食品中猪肉、鸡肉源性成分[J].肉类研究,2016,30(9):17-22.
[16]侯东军,韩合敬,郝智慧,等.三重荧光PCR法鉴定食品中牛、猪、鸭3种动物源性成分[J].畜牧与兽医,2016,48(4):45-48.
[17]曾少灵,秦智锋,阮周曦,等.多重实时荧光PCR检测牛、山羊和绵羊源性成分[J].生物工程学报,2009,25(1):139-146.
[18]黄银花,胡晓湘,徐慰倬,等.影响多重PCR扩增效果的因素[J].遗传,2003,25(1):65-68.
[19]王金凤,王建昌,张伟,等.GNM C7-8实时荧光定量PCR同时快速检测8种食源性致病菌[J].食品安全质量检测学报,2018,9(9):2090-2095.
[20]Q/DXJNM0001-2012 GNM C7-8实时荧光定量PCR分析仪[S].北京:中国标准出版社,2012.
[21]李静,段永生,王建昌,等.实时荧光单引物等温扩增检测阪崎克罗诺杆菌方法的建立[J].中国食品卫生杂志,2015,27(5):524-530.
[22]李瑞,王建昌,李静,等.实时荧光单引物等温扩增(SPIA)技术检测大肠杆菌O157的方法研究[J].现代食品科技,2016,32(2):317-322,316.
[23]朱业培,王玮,吕青骎,等.基于基因芯片技术检测6种动物源性成分[J].南京农业大学学报,2015,38(6):1003-1008.
[24]吴鑫,章志超,王栋,等.基于膜基因芯片技术的肉制品中5种动物源性成分的鉴定[J].现代预防医学,2018,45(10):1762-1765,1786.
[25]Wu Q,Xiang S,Wang W,et al.Species identification of fox-,mink-,dog-,and rabbit-derived ingredients by multiplex PCRand real-time PCR assay[J].Applied Biochemistry&Biotechnology,2017(1):1-12.
[26]Rentsch J,Weibel S,Ruf J,et al.Interlaboratory validation of two multiplex quantitative real-time PCR methods to determine species DNA of cow,sheep and goat as a measure of milk proportions in cheese[J].European Food Research&Technology,2013,236(1):217-227.