ELISA-based detection of fresh and frozen-thawed lamb meat: a promising analytical approach for meat authentication

Fresh/chilled meat differs in quality from frozen-thawed meat, and consumers prefer fresh meat over frozen-thawed meat. Differentiating between the two types of meat is an important part of the meat quality control system. This study aimed to develop an ELISA based on cytoplasmic antioxidant enzyme superoxide dismutase (SOD) as a biomarker. IgGs were raised in experimental animals and purified using immunoaffinity chromatography. The assay was optimized with guinea pig anti-SOD pAb as the capture antibody and rabbit anti-SOD pAb as the detection antibody. The assay showed excellent performance for differentiation, as the ROC area under the curve values were >0.9. A sensitivity of 95.8% and specificity of 95.0% were observed at a positive percentage (PP%) criterion value of 52.752. Sandwich ELISA results showed a significant difference between the chilled and repeatedly frozen-thawed lamb samples. Lamb meat stored for five days in a chiller showed a PP% value below the threshold PP% value, indicating that the assay can differentiate meat until 5 days of chiller storage. In addition, a thawing time of more than 18 h at 4 ± 1 °C is required to differentiate between both types of lamb meat with a size of 500 g. The developed ELISA can be applied at various points in the meat value chain to differentiate fresh meat from frozen-thawed meat.