Establishment of enzyme-linked immunosorbent assay for beef and lamb contents in cooked meat

Abstract

In this study, an enzyme-linked immunosorbent assay (ELISA) was established to detect beef and lamb components, and its performance was tested. Double-antibody sandwich ELISA was adopted and determined a coating concentration of capture antibody 3G5 of 1:4 000, a working concentration of enzyme-labeled antibody 2E7-horseradish peroxidase (HRP) of 1:1 000, a sample incubation time of 60 min and a detection antibody reaction time of 60 min. The specificity, sensitivity, repeatability and stability of this assay were determined. The limit of detection for beef and lamb skeletal muscle troponin I was 45 mg/kg, the inter-assay and intra-assay recovery rates ranged from 80.4% to 115.7%, the coefficients of variation were below 13.6%, and the cross reaction rates of the tissue components of chicken, duck and fish were below 13.4%. The sandwich ELISA method established in this study is stable and has high accuracy. The test results were consistent with the polymerase chain reaction (PCR) method at 50 and 100 g/kg. Therefore, this ELISA method can be used to quantitatively detect beef and lamb components in meat products.