Abstract
Adulteration of cattle meat with other species raises critical public health concerns, as existing detection methods lack sensitivity, specificity and practicality. We address this gap by introducing a novel Point-of-Care (POC) polymerase spiral reaction (PSR) assay. This rapid, on-site tool enables highly specific and sensitive identification of cattle tissue, even in resource-limited settings. This innovation paves the way for improved food safety and consumer protection. The cattle-specific PSR assay employs COX3 gene-targeting primers and a chromogenic indicator triad (HNB, Malachite Green and Phenol Red) for rapid visual detection. Cross-amplification absence with other meats (buffalo, sheep, goat and pig) and concordance with SYBR Green I and agarose gel electrophoresis confirm specificity. PSR at 62 °C for 60minutes exhibits a 100 fg detection limit and identifies 0.1% meat admixing. Efficacy is demonstrated across a diverse range of cattle meat samples, blind samples and processed cattle muscle samples. Compatibility with both conventional (snap chill) and commercial DNA extraction kits enables on-site analysis within 90minutes, facilitating rapid field sample analysis.
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