ArticleFood chemistry. Molecular sciences2026
An ARMS-integrated high-resolution melting assay for reliable discrimination of the A1/A2 polymorphism in the bovine
Article in Food chemistry. Molecular sciences, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Practical, rapid, and cost-efficient screening tools are needed to verify A2 milk purity and support selective breeding programs. Although several genotyping methods are available, their routine implementation is often limited by equipment costs and labor-intensive post-PCR processing. We hypothesized that combining allele-specific primer design with High-Resolution Melting (HRM) analysis would enhance the sensitivity of CSN2 A1/A2 discrimination by generating allele-specific amplicons with more clearly separated melting profiles than conventional HRM, which relies on the subtle melting difference associated with a single nucleotide polymorphism. To test this, we developed an ARMS-integrated HRM assay targeting the CSN2 c.245C > A variant. The assay produced distinct melting patterns that consistently discriminated A1A1, A1A2, and A2A2 genotypes across blood, hair follicle, and milk DNA samples without post-PCR electrophoresis. Application to commercial milk products further demonstrated its potential as a product-level screening tool. In complementary mixture experiments, qPCR showed high linearity (R
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