Rapid Full-Cycle Technique to Control Adulteration of Meat Products: Integration of Accelerated Sample Preparation, Recombinase Polymerase Amplification, and Test-Strip Detection

Verifying the authenticity of food products is essential due to the recent increase in counterfeit meat-containing food products. The existing methods of detection have a number of disadvantages. Therefore, simple, cheap, and sensitive methods for detecting various types of meat are required. In thi...

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Autores principales:	Aleksandr V. Ivanov, Demid S. Popravko, Irina V. Safenkova, Elena A. Zvereva, Boris B. Dzantiev, Anatoly V. Zherdev
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	meat adulteration chicken additives pig additives cytochrome B recombinase polymerase amplification lateral flow assay Organic chemistry QD241-441
Acceso en línea:	https://doaj.org/article/6e318ba694b34212bbc88671dcedccec
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spelling	oai:doaj.org-article:6e318ba694b34212bbc88671dcedccec2021-11-25T18:27:22ZRapid Full-Cycle Technique to Control Adulteration of Meat Products: Integration of Accelerated Sample Preparation, Recombinase Polymerase Amplification, and Test-Strip Detection10.3390/molecules262268041420-3049https://doaj.org/article/6e318ba694b34212bbc88671dcedccec2021-11-01T00:00:00Zhttps://www.mdpi.com/1420-3049/26/22/6804https://doaj.org/toc/1420-3049Verifying the authenticity of food products is essential due to the recent increase in counterfeit meat-containing food products. The existing methods of detection have a number of disadvantages. Therefore, simple, cheap, and sensitive methods for detecting various types of meat are required. In this study, we propose a rapid full-cycle technique to control the chicken or pig adulteration of meat products, including 3 min of crude DNA extraction, 20 min of recombinase polymerase amplification (RPA) at 39 °C, and 10 min of lateral flow assay (LFA) detection. The cytochrome B gene was used in the developed RPA-based test for chicken and pig identification. The selected primers provided specific RPA without DNA nuclease and an additional oligonucleotide probe. As a result, RPA–LFA, based on designed fluorescein- and biotin-labeled primers, detected up to 0.2 pg total DNA per μL, which provided up to 0.001% <i>w</i>/<i>w</i> identification of the target meat component in the composite meat. The RPA–LFA of the chicken and pig meat identification was successfully applied to processed meat products and to meat after heating. The results were confirmed by real-time PCR. Ultimately, the developed analysis is specific and enables the detection of pork and chicken impurities with high accuracy in raw and processed meat mixtures. The proposed rapid full-cycle technique could be adopted for the authentication of other meat products.Aleksandr V. IvanovDemid S. PopravkoIrina V. SafenkovaElena A. ZverevaBoris B. DzantievAnatoly V. ZherdevMDPI AGarticlemeat adulterationchicken additivespig additivescytochrome Brecombinase polymerase amplificationlateral flow assayOrganic chemistryQD241-441ENMolecules, Vol 26, Iss 6804, p 6804 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	meat adulteration chicken additives pig additives cytochrome B recombinase polymerase amplification lateral flow assay Organic chemistry QD241-441
spellingShingle	meat adulteration chicken additives pig additives cytochrome B recombinase polymerase amplification lateral flow assay Organic chemistry QD241-441 Aleksandr V. Ivanov Demid S. Popravko Irina V. Safenkova Elena A. Zvereva Boris B. Dzantiev Anatoly V. Zherdev Rapid Full-Cycle Technique to Control Adulteration of Meat Products: Integration of Accelerated Sample Preparation, Recombinase Polymerase Amplification, and Test-Strip Detection
description	Verifying the authenticity of food products is essential due to the recent increase in counterfeit meat-containing food products. The existing methods of detection have a number of disadvantages. Therefore, simple, cheap, and sensitive methods for detecting various types of meat are required. In this study, we propose a rapid full-cycle technique to control the chicken or pig adulteration of meat products, including 3 min of crude DNA extraction, 20 min of recombinase polymerase amplification (RPA) at 39 °C, and 10 min of lateral flow assay (LFA) detection. The cytochrome B gene was used in the developed RPA-based test for chicken and pig identification. The selected primers provided specific RPA without DNA nuclease and an additional oligonucleotide probe. As a result, RPA–LFA, based on designed fluorescein- and biotin-labeled primers, detected up to 0.2 pg total DNA per μL, which provided up to 0.001% <i>w</i>/<i>w</i> identification of the target meat component in the composite meat. The RPA–LFA of the chicken and pig meat identification was successfully applied to processed meat products and to meat after heating. The results were confirmed by real-time PCR. Ultimately, the developed analysis is specific and enables the detection of pork and chicken impurities with high accuracy in raw and processed meat mixtures. The proposed rapid full-cycle technique could be adopted for the authentication of other meat products.
format	article
author	Aleksandr V. Ivanov Demid S. Popravko Irina V. Safenkova Elena A. Zvereva Boris B. Dzantiev Anatoly V. Zherdev
author_facet	Aleksandr V. Ivanov Demid S. Popravko Irina V. Safenkova Elena A. Zvereva Boris B. Dzantiev Anatoly V. Zherdev
author_sort	Aleksandr V. Ivanov
title	Rapid Full-Cycle Technique to Control Adulteration of Meat Products: Integration of Accelerated Sample Preparation, Recombinase Polymerase Amplification, and Test-Strip Detection
title_short	Rapid Full-Cycle Technique to Control Adulteration of Meat Products: Integration of Accelerated Sample Preparation, Recombinase Polymerase Amplification, and Test-Strip Detection
title_full	Rapid Full-Cycle Technique to Control Adulteration of Meat Products: Integration of Accelerated Sample Preparation, Recombinase Polymerase Amplification, and Test-Strip Detection
title_fullStr	Rapid Full-Cycle Technique to Control Adulteration of Meat Products: Integration of Accelerated Sample Preparation, Recombinase Polymerase Amplification, and Test-Strip Detection
title_full_unstemmed	Rapid Full-Cycle Technique to Control Adulteration of Meat Products: Integration of Accelerated Sample Preparation, Recombinase Polymerase Amplification, and Test-Strip Detection
title_sort	rapid full-cycle technique to control adulteration of meat products: integration of accelerated sample preparation, recombinase polymerase amplification, and test-strip detection
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/6e318ba694b34212bbc88671dcedccec
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Rapid Full-Cycle Technique to Control Adulteration of Meat Products: Integration of Accelerated Sample Preparation, Recombinase Polymerase Amplification, and Test-Strip Detection

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