一种食品中猪源性成分的快速检测技术
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摘要
建立一种基于PCR技术的食品中猪源性成分快速检测技术。根据猪线粒体cytb基因设计引物,进行PCR扩增。该方法对猪DNA检测的灵敏度达到100pg;该方法仅对猪DNA检测呈阳性,与其他物种DNA无交叉反应;同时可用于商业样本及复杂食物样本的检测。本研究的猪源性成分特异PCR检测技术具有快速、准确的特点,且具有较高的特异性和敏感性,可作为肉制品中猪源性成分快速检测的手段。
To develop a fast identification technology based on PCR for the detection of pig ingredient in meat products,primers were designed according to the mitochondrial cytochrome-b sequences of pig, then amplified by PCR. The limit of detection was up to 100 pg of pig DNA; the test was only specific for pig, and no cross-reactions were observed in other non-target species. Meanwhile, it can be used for commercial and complex food samples. In this study, the pig ingredient specific PCR detection technology has the characteristic of rapidly and accurately, and with higher specificity and sensitivity, it is a rapid approach to detect pig ingredient in meat products.
To develop a fast identification technology based on PCR for the detection of pig ingredient in meat products,primers were designed according to the mitochondrial cytochrome-b sequences of pig, then amplified by PCR. The limit of detection was up to 100 pg of pig DNA; the test was only specific for pig, and no cross-reactions were observed in other non-target species. Meanwhile, it can be used for commercial and complex food samples. In this study, the pig ingredient specific PCR detection technology has the characteristic of rapidly and accurately, and with higher specificity and sensitivity, it is a rapid approach to detect pig ingredient in meat products.
引文
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[12]NADIA H,IMAD N,AL-SAFADI B.Identification of meat species by PCR-RFLP of the mitochondrial COI gene[J].Meat Science,2011(90):490-493.
[13]Karabasanavar,N.S.,Singh,S.P.,Umapathi,V.,Kumar,D.,Patil,G.,&Shebannavar,S.N.(2011).A highly specific PCRassay for identification of raw and heat treated mutton(Ovis aries).Small Ruminant Research,100(2),153-158.
[14]薛超波,王萍亚,李素芳,等.基于PCR多物种鉴定技术及其在肉类鉴定中的应用[J].食品安全质量检测学报,2016,7(2):562-566.
[2]王瑞元,巢强国,葛宇,等.奶糖中猪成分明胶PCR鉴定方法的研究[J].食品工业,2010(1):86-88.
[3]李巧玲,刘景艳.市场鲜猪肉掺假状况的调查监测[J].食品科学,2004,25(10):273-276.
[4]罗家琴,王加启,卜登攀,等.饲料中牛,羊,猪,鸡源性成分的PCR检测方法及其应用[J].中国农业科学,2008,41(7):2112-2119.
[5]李家鹏,乔晓玲,田寒友,等.食品和饲料中动物源性成分检测技术研究进展[J].食品科学.
[6]Renon,P.,Bernardi,C.,Scocca,S.,Cantoni,C.,Gridavilla,G.,2003.IEF(Iso-electricfocussing)and gas chromatography to identi2011,32(9):340-347.fy wild ruminant species.Indus.Aliment.42,496-500.
[7]Ashoor,S.H.,Monte,W.C.,&Stiles,P.G.(1998).Liquid chromatographic identification of meats.Journal-Association of Official Analytical Chemists,71(2),397-403.
[8]Hajmeer,M.,Cliiver,D.O.,&Provost,R.(2003).Spinal cord tissue detection in comminuted beef:comparison of two immunological methods.Meat Sciences,31(3),159-163.
[9]ARSLAN A,ILHAK O I,CALICIOGLU M.Effect of method of cooking on identification of heat processed beef using polymerase chain reaction(PCR)technique[J].Meat Science,2006,72(2):326-330.DOI:10.1016/j.meatsci.2005.08.001.
[10]UNSELD M,BEYERMANN B,BRANDT P,et al.I-denti?cation of the species origin of highly processed meat products by mitochondrial DNA sequences[J].PCR Methods and Applications,1995,4:241-243.
[11]Chikuni,K.,Ozustume,K.,Hoishikawa,T.,&Kato,S.(1990).Species identification of cooked meats by DNA hybridization assay.Meat Science,27(2),199-208.
[12]NADIA H,IMAD N,AL-SAFADI B.Identification of meat species by PCR-RFLP of the mitochondrial COI gene[J].Meat Science,2011(90):490-493.
[13]Karabasanavar,N.S.,Singh,S.P.,Umapathi,V.,Kumar,D.,Patil,G.,&Shebannavar,S.N.(2011).A highly specific PCRassay for identification of raw and heat treated mutton(Ovis aries).Small Ruminant Research,100(2),153-158.
[14]薛超波,王萍亚,李素芳,等.基于PCR多物种鉴定技术及其在肉类鉴定中的应用[J].食品安全质量检测学报,2016,7(2):562-566.