Western blot detection of brain material in heated meat products using myelin basic protein and neuron-specific enolase as biomarkers

摘要

An assay based on Western blotting and detection of central nervous system (CNS)-specific antigens was developed to detect brain tissue in processed (heated) meat products.Bands of antigen-bound primary antibodies were visualised through secondary anti-antibodies labelled with peroxidase,which generated chemiluminescence documented by a photographic film.Ponceau-S staining before antibody incubation and molecular mass information on detected antigens after immunoreactions added information supporting correct identification of brain tissue in the meat products.In this approach B50/growth-associated protein (B50),glial fibrillary acidic protein (GFAP),myelin basic protein (MBP),neurofilament (NF),neuron-specific enolase (NSE) and synaptophysin (Syn) proteins were detected in raw luncheon meat and a liver product enriched with brain tissue at a level of 5% (m/m).Only MBP and NSE were considered suitable biomarkers for detection of 1% (m/m) brain tissue in meat products pasteurised at 70 deg C or sterilised at 115 deg C.The use of an anti-monkey MBP instead of anti-human MBP enabled speciation of the CNS material whether from bovine and ovine brains or from porcine brain tissue.This immunoblot assay potentiates the analysis of approximately 70 samples within 8 h,including sample preparation and the simultaneous probing of NSE and MBP target antigens.