Validation of a fast real-time PCR method to detect fraud and mislabeling in milk and dairy products
Fast real-time PCR TaqMan assays were developed and validated for species identification in dairy products. Based on the amplification of 12S rRNA and cytB partial genes of mitochondrial DNA, the methods were demonstrated to be sensitive, fast, and species-specific for Bos taurus, Ovis aries, Bubalu...
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| Veröffentlicht in: | Journal of dairy science 2017-01, Vol.100 (1), p.106-112 |
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| container_title | Journal of dairy science |
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| creator | Di Domenico, M. Di Giuseppe, M. Wicochea Rodríguez, J.D. Cammà, C. |
| description | Fast real-time PCR TaqMan assays were developed and validated for species identification in dairy products. Based on the amplification of 12S rRNA and cytB partial genes of mitochondrial DNA, the methods were demonstrated to be sensitive, fast, and species-specific for Bos taurus, Ovis aries, Bubalus bubalis, and Capra hircus. The limit of detection calculated was lower than 1%, and the efficiency was reported to be higher than 96% in every assay. An internal amplification control was used to detect possible false negatives. The method was validated by means of laboratory-prepared samples mixing different species. Moreover, 18 commercial dairy samples were analyzed by both real-time PCR and isoelectric focusing, the official European Union reference method. The 4 TaqMan assays were confirmed to be a useful tool for milk and dairy product authentication. |
| doi_str_mv | 10.3168/jds.2016-11695 |
| format | Article |
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Based on the amplification of 12S rRNA and cytB partial genes of mitochondrial DNA, the methods were demonstrated to be sensitive, fast, and species-specific for Bos taurus, Ovis aries, Bubalus bubalis, and Capra hircus. The limit of detection calculated was lower than 1%, and the efficiency was reported to be higher than 96% in every assay. An internal amplification control was used to detect possible false negatives. The method was validated by means of laboratory-prepared samples mixing different species. Moreover, 18 commercial dairy samples were analyzed by both real-time PCR and isoelectric focusing, the official European Union reference method. 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Based on the amplification of 12S rRNA and cytB partial genes of mitochondrial DNA, the methods were demonstrated to be sensitive, fast, and species-specific for Bos taurus, Ovis aries, Bubalus bubalis, and Capra hircus. The limit of detection calculated was lower than 1%, and the efficiency was reported to be higher than 96% in every assay. An internal amplification control was used to detect possible false negatives. The method was validated by means of laboratory-prepared samples mixing different species. Moreover, 18 commercial dairy samples were analyzed by both real-time PCR and isoelectric focusing, the official European Union reference method. The 4 TaqMan assays were confirmed to be a useful tool for milk and dairy product authentication.</description><subject>Animals</subject><subject>Buffaloes</subject><subject>Cattle</subject><subject>dairy products</subject><subject>Dairy Products - analysis</subject><subject>Food Analysis - methods</subject><subject>Goats</subject><subject>isoelectric focusing</subject><subject>Limit of Detection</subject><subject>Milk - chemistry</subject><subject>real-time PCR</subject><subject>Real-Time Polymerase Chain Reaction - methods</subject><subject>Sensitivity and Specificity</subject><subject>Sheep, Domestic</subject><subject>species identification</subject><subject>Species Specificity</subject><issn>0022-0302</issn><issn>1525-3198</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kMtPAyEQh4nR2Pq4ejQcvWwFdqHs0TS-kiYao14JC4NS91GBNfG_F1v15okM-eY3Mx9CJ5TMSirk-crGGSNUFJSKmu-gKeWMFyWt5S6aEsJYQUrCJuggxlUuKSN8H03YXArOSTVF9lm33urkhx4PDmvsdEw4gG6L5DvA94sH3EF6HSxOA7aQwCTsgh4t1r3FnY-tbqD1_Qv2fS7bt82_1T584nUY7GhSPEJ7TrcRjn_eQ_R0dfm4uCmWd9e3i4tlYUpZpYJToee15bSSRDSUlY12dUnknIDL2wrpTEm1sNXcMeZqcA0QaRwXlJCKcFkeorNtbh78PkJMKu9noG11D8MYFZUV45UQvMrobIuaMMQYwKl18J0On4oS9W1WZbPq26zamM0Npz_ZY9OB_cN_VWZAbgHIF354CCoaD70B60OWpuzg_8v-AhgOhhk</recordid><startdate>201701</startdate><enddate>201701</enddate><creator>Di Domenico, M.</creator><creator>Di Giuseppe, M.</creator><creator>Wicochea Rodríguez, J.D.</creator><creator>Cammà, C.</creator><general>Elsevier Inc</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>201701</creationdate><title>Validation of a fast real-time PCR method to detect fraud and mislabeling in milk and dairy products</title><author>Di Domenico, M. ; Di Giuseppe, M. ; Wicochea Rodríguez, J.D. ; Cammà, C.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c384t-516a79d514806b123baf930870ef55068fc31a6d47f22f9efbe08cf5610040583</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Animals</topic><topic>Buffaloes</topic><topic>Cattle</topic><topic>dairy products</topic><topic>Dairy Products - analysis</topic><topic>Food Analysis - methods</topic><topic>Goats</topic><topic>isoelectric focusing</topic><topic>Limit of Detection</topic><topic>Milk - chemistry</topic><topic>real-time PCR</topic><topic>Real-Time Polymerase Chain Reaction - methods</topic><topic>Sensitivity and Specificity</topic><topic>Sheep, Domestic</topic><topic>species identification</topic><topic>Species Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Di Domenico, M.</creatorcontrib><creatorcontrib>Di Giuseppe, M.</creatorcontrib><creatorcontrib>Wicochea Rodríguez, J.D.</creatorcontrib><creatorcontrib>Cammà, C.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of dairy science</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Di Domenico, M.</au><au>Di Giuseppe, M.</au><au>Wicochea Rodríguez, J.D.</au><au>Cammà, C.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Validation of a fast real-time PCR method to detect fraud and mislabeling in milk and dairy products</atitle><jtitle>Journal of dairy science</jtitle><addtitle>J Dairy Sci</addtitle><date>2017-01</date><risdate>2017</risdate><volume>100</volume><issue>1</issue><spage>106</spage><epage>112</epage><pages>106-112</pages><issn>0022-0302</issn><eissn>1525-3198</eissn><abstract>Fast real-time PCR TaqMan assays were developed and validated for species identification in dairy products. 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| source | MEDLINE; Elsevier ScienceDirect Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals |
| subjects | Animals Buffaloes Cattle dairy products Dairy Products - analysis Food Analysis - methods Goats isoelectric focusing Limit of Detection Milk - chemistry real-time PCR Real-Time Polymerase Chain Reaction - methods Sensitivity and Specificity Sheep, Domestic species identification Species Specificity |
| title | Validation of a fast real-time PCR method to detect fraud and mislabeling in milk and dairy products |
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