肉制品中猪源性成分相对定量检测方法研究
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摘要
目的建立一种肉制品中猪源性成分的相对定量检测方法。方法针对猪的ER-beta基因的mRNA序列设计了特异性引物与探针序列,并对引物和探针的特异性进行了验证。构建重组质粒,利用质粒拷贝数的log值和Ct值构建标准曲线。通过标准曲线计算各个样本拷贝数,将待测样本和标准种源样本的拷贝数进行比较,确定检测样本的猪源性含量。结果实际掺加量分别为20%、40%、50%和60%的混合样品经该方法检测所得掺假量均值为猪体系24.8%、53.1%、53.1%、61%,检测结果与其实际含量基本一致。结论该方法基本能实现肉制品中猪源性成分的相对定量检测。
Objective To establish a relative quantitative method for the detection of porcine-derived components in meat products. Methods Specific primers and probe sequences were designed for porcine ER-beta-based mRNA gene sequence, and the specificity of the primers and probes was verified. A recombinant plasmid was constructed, and a standard curve was constructed by the log value and Ct value of the plasmid copy number. The copy number of each sample was calculated by the standard curve, the copy number of the sample was compared to be tested with the standard provenance sample, and the porcine-derived content of the test sample was determined. Results The average adulteration of the mixed samples with actual addition amounts of 20%, 40%,50% and 60% by the method were 24.8%, 53.1%, 53.1%, 61% of the pig system. The test results were basically the same as their actual content. Conclusion This method basically can realize the relative quantitative detection of the porcine-derived components in the meat product.
Objective To establish a relative quantitative method for the detection of porcine-derived components in meat products. Methods Specific primers and probe sequences were designed for porcine ER-beta-based mRNA gene sequence, and the specificity of the primers and probes was verified. A recombinant plasmid was constructed, and a standard curve was constructed by the log value and Ct value of the plasmid copy number. The copy number of each sample was calculated by the standard curve, the copy number of the sample was compared to be tested with the standard provenance sample, and the porcine-derived content of the test sample was determined. Results The average adulteration of the mixed samples with actual addition amounts of 20%, 40%,50% and 60% by the method were 24.8%, 53.1%, 53.1%, 61% of the pig system. The test results were basically the same as their actual content. Conclusion This method basically can realize the relative quantitative detection of the porcine-derived components in the meat product.
引文
[1]刘秀梅.中国城乡居民动物性食物消费研究[D].北京:中国农业大学,2005.Liu XM.Empirical comparison of animal food consumption between urban and rural residents in China[D].Beijing:China Agricultural University,2005.
[2]范冰洁.肉制品中动物源性成分检测方法研究进展[J].新疆畜牧业,2018,(3):10-12,16Fan BJ.Advances in methods for determination of animal-derived components in meat products[J].J Xinjiang Anim Husbandr,2018,(3):10-12,16.
[3]冯秀燕,董红霞,魏秀莲,等.动物源性饲料中牛源和羊源性成分的检测方法[J].饲料研究,2006,(11):52-54.Feng XX,Dong HX,Wei XL,et al.Methods for the determination of bovine and sheep derived components in animal-derived feed[J].Feed Res,2006,(11):52-54.
[4]卜登攀,王加启,贺云霞,等.动物源性饲料检测技术研究进展[J].中国畜牧兽医,2008,35(2):54-59.Bo DP,Wang JQ,He YX,et al.Detection of the animal origin materials in feedstuff:A review[J].China Anim Husbandr Veter Med,2008,35(2):54-59.
[5]杨冬燕,李浩,杨永存.肉制品掺假鉴别与定量检测[J].卫生研究,2016,45(2):324-328.Yang DY,Li H,Yang YC.Identification and quantitative detection of adulterated meat products[J].J Hyg Res,2016,45(2):324-328.
[6]Yin RH,Bai WL,Wang JM,et al.Development of an assay for rapid identification ofmeat from yak and cattle using polymerase chainreaction technique[J].Meat Sci,2009,83(1):38-44.
[7]Ghovvati S,Nassiri MR,Mirhoseini SZ,et al.Fraud identification in industrial meat productsby multiplex PCR assay[J].Food Contr,2009,20(8):696-699.
[8]杨冬燕,韦梅霞,杨永存,等.多重荧光PCR鉴别羊肉掺假[J].食品安全质量检测学报,2015,6(2):555-562.Yang DY,Wei MX,Yang YC,et al.Identification of mutton adulteration by multiplex fluorescent PCR[J].J Food Saf Qual,2015,6(2):555-562.
[9]Doosti A,Ghasemi DP,Rahimi E.Molecular assay to fraud identification of meat products[J].J Food Sci Technol,2014,51(1):148-152.
[10]Rastogi C,Dhame MS,Walujkar S,et al.Species identification and authentication of tissues of animal origin using mitochondial and nuclear markers[J].Meat Sci,2007,(76):666-674.
[11]刘岑杰,刘彦泓,杨滴,等.肉制品中鸭源性成分的实时荧光PCR检测[J].肉类工业,2015,(1):1,51-53.Liu CJ,Liu YH,Yang D,et al.Detection of components of duck origin in meat products with real-time PCR method[J].Meat Ind,2015,(1):1,51-53.
[12]张驰,邱皓璞,张筠.荧光定量PCR检测肉制品中鸭源性成分[J].食品科学,2013,34(18):154-157.Zhang C,Qiu HP,Zhang Y.A quantitative fluorescent PCR method for detection of duck-derived ingredients in meat products[J]J Food Sci,2013,34(18):154-157.
[13]Laube I,Zagon J,Hermann B.Quantitative determination of commercially relevant species in foods by real-time PCR[J].Inter J Food Sci Technol,2007,42(3):336-341.
[14]曹建民,田露,张越杰.我国牛肉消费及其对猪肉价格变化的反应研究[J].中国畜牧杂志,2012,48(12):12-15.Cao JM,Tian L,Zhang YJ.Research on beef consumption and its response to the change of pork price in China[J].Chin J Anim Husbandr,2012,48(12):12-15.
[15]王贝贝,肖海峰.中国羊肉供需状况及展望[J].农业展望,2012,(10):19-22.Wang BB,Xiao HF.Supply and demand situation and prospect of mutton in China[J].Agric Outlook,2012,(10):19-22.
[2]范冰洁.肉制品中动物源性成分检测方法研究进展[J].新疆畜牧业,2018,(3):10-12,16Fan BJ.Advances in methods for determination of animal-derived components in meat products[J].J Xinjiang Anim Husbandr,2018,(3):10-12,16.
[3]冯秀燕,董红霞,魏秀莲,等.动物源性饲料中牛源和羊源性成分的检测方法[J].饲料研究,2006,(11):52-54.Feng XX,Dong HX,Wei XL,et al.Methods for the determination of bovine and sheep derived components in animal-derived feed[J].Feed Res,2006,(11):52-54.
[4]卜登攀,王加启,贺云霞,等.动物源性饲料检测技术研究进展[J].中国畜牧兽医,2008,35(2):54-59.Bo DP,Wang JQ,He YX,et al.Detection of the animal origin materials in feedstuff:A review[J].China Anim Husbandr Veter Med,2008,35(2):54-59.
[5]杨冬燕,李浩,杨永存.肉制品掺假鉴别与定量检测[J].卫生研究,2016,45(2):324-328.Yang DY,Li H,Yang YC.Identification and quantitative detection of adulterated meat products[J].J Hyg Res,2016,45(2):324-328.
[6]Yin RH,Bai WL,Wang JM,et al.Development of an assay for rapid identification ofmeat from yak and cattle using polymerase chainreaction technique[J].Meat Sci,2009,83(1):38-44.
[7]Ghovvati S,Nassiri MR,Mirhoseini SZ,et al.Fraud identification in industrial meat productsby multiplex PCR assay[J].Food Contr,2009,20(8):696-699.
[8]杨冬燕,韦梅霞,杨永存,等.多重荧光PCR鉴别羊肉掺假[J].食品安全质量检测学报,2015,6(2):555-562.Yang DY,Wei MX,Yang YC,et al.Identification of mutton adulteration by multiplex fluorescent PCR[J].J Food Saf Qual,2015,6(2):555-562.
[9]Doosti A,Ghasemi DP,Rahimi E.Molecular assay to fraud identification of meat products[J].J Food Sci Technol,2014,51(1):148-152.
[10]Rastogi C,Dhame MS,Walujkar S,et al.Species identification and authentication of tissues of animal origin using mitochondial and nuclear markers[J].Meat Sci,2007,(76):666-674.
[11]刘岑杰,刘彦泓,杨滴,等.肉制品中鸭源性成分的实时荧光PCR检测[J].肉类工业,2015,(1):1,51-53.Liu CJ,Liu YH,Yang D,et al.Detection of components of duck origin in meat products with real-time PCR method[J].Meat Ind,2015,(1):1,51-53.
[12]张驰,邱皓璞,张筠.荧光定量PCR检测肉制品中鸭源性成分[J].食品科学,2013,34(18):154-157.Zhang C,Qiu HP,Zhang Y.A quantitative fluorescent PCR method for detection of duck-derived ingredients in meat products[J]J Food Sci,2013,34(18):154-157.
[13]Laube I,Zagon J,Hermann B.Quantitative determination of commercially relevant species in foods by real-time PCR[J].Inter J Food Sci Technol,2007,42(3):336-341.
[14]曹建民,田露,张越杰.我国牛肉消费及其对猪肉价格变化的反应研究[J].中国畜牧杂志,2012,48(12):12-15.Cao JM,Tian L,Zhang YJ.Research on beef consumption and its response to the change of pork price in China[J].Chin J Anim Husbandr,2012,48(12):12-15.
[15]王贝贝,肖海峰.中国羊肉供需状况及展望[J].农业展望,2012,(10):19-22.Wang BB,Xiao HF.Supply and demand situation and prospect of mutton in China[J].Agric Outlook,2012,(10):19-22.