ArticleCurrent developments in nutrition2026
A Rapid Genotyping Assay for
Article in Current developments in nutrition, 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
Background: Nutrigenetic research linking fat mass and obesity-associated ( Objectives: The aim of this study is to develop, validate, and apply a cost-effective 4-plex high-resolution melting (HRM) assay for simultaneous detection of Methods: Allele-specific primers with distinct melting temperatures were designed for multiplexed HRM detection of 4 obesity-associated single nucleotide polymorphisms. Analytical validation was performed against Sanger sequencing in 30 samples. Diagnostic accuracy was assessed by sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). The validated assay was applied to genotype 384 Thai adults (123 normal-weight, 261 with overweight/obesity) from Bangkok, Thailand. Genotype frequencies and Hardy-Weinberg equilibrium (HWE) were evaluated by the χ Results: The 4-plex HRM assay demonstrated 100% concordance with Sanger sequencing across all 4 variants (sensitivity: 100%; specificity: 100%; PPV: 100%; NPV: 100%). All triplicate measurements produced consistent genotype calls. Total assay time was 70 min. Based on the manufacturer's catalog prices (QIAGEN, 2024), the estimated reagent cost per sample was ∼4- to 5-fold lower than 4 individual TaqMan assays and 6- to 8-fold lower than Sanger sequencing for the same 4 targets. Population screening revealed minor allele frequencies of 5.3%-8.2% for Conclusions: This validated 4-plex HRM assay provides a rapid, accurate, and cost-effective tool for nutrigenetic screening of obesity-associated variants. The method is suitable for large-scale population studies where budget and infrastructure constraints limit access to high-throughput genotyping platforms.
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