Comparative Analysis of Rapid and Less Invasive Methods for A2A2 Dairy Cattle Genotyping and A2 Milk Purity Detection

In this study, two methods for assessing the purity of A2 fermented milk and β-casein genotypes in dairy cows were examined. The need for rapid and precise methods for herd screening and A2 milk quality control justified this study. Accordingly, an ELISA test was developed to identify β-casein A1 in...

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Main Authors: Leandra Oliveira Xavier Albiero, Rafaela Ansiliero, Aniela Pinto Kempka
Format: Article
Language:English
Published: MDPI AG 2024-12-01
Series:Dairy
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Online Access:https://www.mdpi.com/2624-862X/5/4/57
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author Leandra Oliveira Xavier Albiero
Rafaela Ansiliero
Aniela Pinto Kempka
author_facet Leandra Oliveira Xavier Albiero
Rafaela Ansiliero
Aniela Pinto Kempka
author_sort Leandra Oliveira Xavier Albiero
collection DOAJ
description In this study, two methods for assessing the purity of A2 fermented milk and β-casein genotypes in dairy cows were examined. The need for rapid and precise methods for herd screening and A2 milk quality control justified this study. Accordingly, an ELISA test was developed to identify β-casein A1 in fermented milk, and a commercial Lateral Flow Immunoassay (LFIA) was evaluated to determine A2A2 genotypes and the purity of A2 milk. The results demonstrated 100% sensitivity and specificity of the ELISA test in identifying β-casein A1. The LFIA test successfully identified A2A2 genotypes and confirmed the purity of A2 milk, with a minimum detectable contamination of 5% for raw milk and 10% for fermented milk. Both tests exhibited 100% sensitivity and specificity, resulting in positive and negative predictive values of 100%. The positive likelihood ratio was infinite, while the negative was zero, indicating a precise and reliable test with no false diagnoses. Compared to traditional genotyping, these methods proved to be more practical and showed potential for large-scale screening. It was concluded that ELISA and LFIA are valuable tools for ensuring the quality and authenticity of A2 milk, meeting the demands of producers and consumers for safe and healthy dairy products.
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spelling doaj-art-45097e451f8248ce9cd513e84d75135d2025-08-20T02:00:41ZengMDPI AGDairy2624-862X2024-12-015478679910.3390/dairy5040057Comparative Analysis of Rapid and Less Invasive Methods for A2A2 Dairy Cattle Genotyping and A2 Milk Purity DetectionLeandra Oliveira Xavier Albiero0Rafaela Ansiliero1Aniela Pinto Kempka2Multicenter Program in Postgraduate in Biochemistry and Molecular Biology, Department of Animal Production and Food, Santa Catarina State University, Lages 88520-000, BrazilMulticenter Program in Postgraduate in Biochemistry and Molecular Biology, Department of Animal Production and Food, Santa Catarina State University, Lages 88520-000, BrazilMulticenter Program in Postgraduate in Biochemistry and Molecular Biology, Department of Food Engineering and Chemical Engineering, Santa Catarina State University, Pinhalzinho 89870-000, BrazilIn this study, two methods for assessing the purity of A2 fermented milk and β-casein genotypes in dairy cows were examined. The need for rapid and precise methods for herd screening and A2 milk quality control justified this study. Accordingly, an ELISA test was developed to identify β-casein A1 in fermented milk, and a commercial Lateral Flow Immunoassay (LFIA) was evaluated to determine A2A2 genotypes and the purity of A2 milk. The results demonstrated 100% sensitivity and specificity of the ELISA test in identifying β-casein A1. The LFIA test successfully identified A2A2 genotypes and confirmed the purity of A2 milk, with a minimum detectable contamination of 5% for raw milk and 10% for fermented milk. Both tests exhibited 100% sensitivity and specificity, resulting in positive and negative predictive values of 100%. The positive likelihood ratio was infinite, while the negative was zero, indicating a precise and reliable test with no false diagnoses. Compared to traditional genotyping, these methods proved to be more practical and showed potential for large-scale screening. It was concluded that ELISA and LFIA are valuable tools for ensuring the quality and authenticity of A2 milk, meeting the demands of producers and consumers for safe and healthy dairy products.https://www.mdpi.com/2624-862X/5/4/57A2A2 dairy cowsgenotypingELISALFIAcomparison of methodsfacilitated analysis
spellingShingle Leandra Oliveira Xavier Albiero
Rafaela Ansiliero
Aniela Pinto Kempka
Comparative Analysis of Rapid and Less Invasive Methods for A2A2 Dairy Cattle Genotyping and A2 Milk Purity Detection
Dairy
A2A2 dairy cows
genotyping
ELISA
LFIA
comparison of methods
facilitated analysis
title Comparative Analysis of Rapid and Less Invasive Methods for A2A2 Dairy Cattle Genotyping and A2 Milk Purity Detection
title_full Comparative Analysis of Rapid and Less Invasive Methods for A2A2 Dairy Cattle Genotyping and A2 Milk Purity Detection
title_fullStr Comparative Analysis of Rapid and Less Invasive Methods for A2A2 Dairy Cattle Genotyping and A2 Milk Purity Detection
title_full_unstemmed Comparative Analysis of Rapid and Less Invasive Methods for A2A2 Dairy Cattle Genotyping and A2 Milk Purity Detection
title_short Comparative Analysis of Rapid and Less Invasive Methods for A2A2 Dairy Cattle Genotyping and A2 Milk Purity Detection
title_sort comparative analysis of rapid and less invasive methods for a2a2 dairy cattle genotyping and a2 milk purity detection
topic A2A2 dairy cows
genotyping
ELISA
LFIA
comparison of methods
facilitated analysis
url https://www.mdpi.com/2624-862X/5/4/57
work_keys_str_mv AT leandraoliveiraxavieralbiero comparativeanalysisofrapidandlessinvasivemethodsfora2a2dairycattlegenotypinganda2milkpuritydetection
AT rafaelaansiliero comparativeanalysisofrapidandlessinvasivemethodsfora2a2dairycattlegenotypinganda2milkpuritydetection
AT anielapintokempka comparativeanalysisofrapidandlessinvasivemethodsfora2a2dairycattlegenotypinganda2milkpuritydetection