Sika deer (Cervus nippon)-specific real-time PCR method to detect fraudulent labelling of meat and meat products

Abstract Since game meat is more valuable and expensive than meat from domesticated animal species it is a potential target for adulteration. Analytical methods must allow the identification and quantification of meat species to be applicable for the detection of fraudulent labelling. We developed a...

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Autores principales:	Maria Kaltenbrunner, Rupert Hochegger, Margit Cichna-Markl
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/17b0c00957f0477d852ecb356d2fb8cc
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spelling	oai:doaj.org-article:17b0c00957f0477d852ecb356d2fb8cc2021-12-02T11:40:46ZSika deer (Cervus nippon)-specific real-time PCR method to detect fraudulent labelling of meat and meat products10.1038/s41598-018-25299-72045-2322https://doaj.org/article/17b0c00957f0477d852ecb356d2fb8cc2018-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-25299-7https://doaj.org/toc/2045-2322Abstract Since game meat is more valuable and expensive than meat from domesticated animal species it is a potential target for adulteration. Analytical methods must allow the identification and quantification of meat species to be applicable for the detection of fraudulent labelling. We developed a real-time PCR assay for the authentication of sika deer (Cervus nippon) and products thereof. The primer/probe system amplifies a 71 bp fragment of the kappa-casein precursor gene. Since the target sequence contained only one sika deer-specific base, we introduced a deliberate base mismatch in the forward primer. The real-time PCR assay did not show cross-reactivity with 19 animal and 49 plant species tested. Low cross-reactivity was observed with red deer, fallow deer, reindeer and moose. However, with a ΔCt value of ≥11.79 between sika deer and the cross-reacting species, cross-reactivity will not affect the accuracy of the method. LOD and LOQ, determined by analysing serial dilutions of a DNA extract containing 1% (w/w) sika deer DNA in pig DNA, were 0.3% and 0.5%, respectively. The accuracy was evaluated by analysing DNA mixtures and DNA isolates from meat extract mixtures and meat mixtures. In general, recoveries were in the range from 70 to 130%.Maria KaltenbrunnerRupert HocheggerMargit Cichna-MarklNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-11 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Maria Kaltenbrunner Rupert Hochegger Margit Cichna-Markl Sika deer (Cervus nippon)-specific real-time PCR method to detect fraudulent labelling of meat and meat products
description	Abstract Since game meat is more valuable and expensive than meat from domesticated animal species it is a potential target for adulteration. Analytical methods must allow the identification and quantification of meat species to be applicable for the detection of fraudulent labelling. We developed a real-time PCR assay for the authentication of sika deer (Cervus nippon) and products thereof. The primer/probe system amplifies a 71 bp fragment of the kappa-casein precursor gene. Since the target sequence contained only one sika deer-specific base, we introduced a deliberate base mismatch in the forward primer. The real-time PCR assay did not show cross-reactivity with 19 animal and 49 plant species tested. Low cross-reactivity was observed with red deer, fallow deer, reindeer and moose. However, with a ΔCt value of ≥11.79 between sika deer and the cross-reacting species, cross-reactivity will not affect the accuracy of the method. LOD and LOQ, determined by analysing serial dilutions of a DNA extract containing 1% (w/w) sika deer DNA in pig DNA, were 0.3% and 0.5%, respectively. The accuracy was evaluated by analysing DNA mixtures and DNA isolates from meat extract mixtures and meat mixtures. In general, recoveries were in the range from 70 to 130%.
format	article
author	Maria Kaltenbrunner Rupert Hochegger Margit Cichna-Markl
author_facet	Maria Kaltenbrunner Rupert Hochegger Margit Cichna-Markl
author_sort	Maria Kaltenbrunner
title	Sika deer (Cervus nippon)-specific real-time PCR method to detect fraudulent labelling of meat and meat products
title_short	Sika deer (Cervus nippon)-specific real-time PCR method to detect fraudulent labelling of meat and meat products
title_full	Sika deer (Cervus nippon)-specific real-time PCR method to detect fraudulent labelling of meat and meat products
title_fullStr	Sika deer (Cervus nippon)-specific real-time PCR method to detect fraudulent labelling of meat and meat products
title_full_unstemmed	Sika deer (Cervus nippon)-specific real-time PCR method to detect fraudulent labelling of meat and meat products
title_sort	sika deer (cervus nippon)-specific real-time pcr method to detect fraudulent labelling of meat and meat products
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/17b0c00957f0477d852ecb356d2fb8cc
work_keys_str_mv	AT mariakaltenbrunner sikadeercervusnipponspecificrealtimepcrmethodtodetectfraudulentlabellingofmeatandmeatproducts AT ruperthochegger sikadeercervusnipponspecificrealtimepcrmethodtodetectfraudulentlabellingofmeatandmeatproducts AT margitcichnamarkl sikadeercervusnipponspecificrealtimepcrmethodtodetectfraudulentlabellingofmeatandmeatproducts
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Sika deer (Cervus nippon)-specific real-time PCR method to detect fraudulent labelling of meat and meat products

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