Establishment of enzyme-linked immunosorbent assay for beef and lamb contents in cooked meat
Yujing Li, Jingjing Liu, Sufang Fan, Li Zhao, Jing Zhang, Erjing Zhang, Ziran Li, Yan Zhang, Chunsheng Li
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.