Real-time monitoring of cattle (Bos taurus) tissue using a novel point-of-care (POC) polymerase spiral reaction (PSR) colorimetric assay

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 60 minutes 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 90 minutes, facilitating rapid field sample analysis. [Display omitted] •A novel PSR-based POC diagnostic tool for rapid cattle tissue identification.•On-site visual readout employing HNB, Malachite Green, and Phenol Red indicators.•PSR boasts 100 % specificity and 100 fg sensitivity, detecting 0.1 % meat admixing.•PSR facilitates rapid cattle meat identification, completed in less than 90 minutes.