Detection of Pork Fat in Heat-processed Beef Meat Products by ELISA Using Monoclonal Antibody Specific to Pork Fat Protein

Introduction:  Meat and food products containing pork meat and fat may cause serious matter for a Muslim, Jewish and for vegetarians. Therefore, detection and identification of pork adulterated in meat products and food is concerned as an urgent need.

Purpose:  The objective of this study was to develop and optimize an indirect ELISA using anti-pork fat monoclonal antibody (MAb) for the rapid detection of pork fat and its application to the detection of pork fat in heat-processed beef meat products such as autoclaved, steamed, roasted and fried meat.

Methods:  To improve the sensitivity of the indirect ELISA, the optimal sample pre-cooking time, sample coating time and temperature, primary and secondary antibody dilution time were tested. Various buffer systems including sample dilution buffer and blocking buffer were optimized. Heat processed beef meat samples artificially adulterated with known amounts of pork fat (0, 1, 5, 10, 15, 30, and 100%, w/w) were prepared by autoclaving, steaming, roasting and frying and analyzed by the indirect ELISA.

Results:  A standard curve was obtained based on the optimized conditions and the developed indirect ELISA method can sensitively detect 1 % pork fat in heat-processed (autoclaving, steaming, roasting and frying) beef meat. No cross-reaction to other meats such as beef, chicken, goat, duck, horse, and sheep was not obtained in the direct ELISA system.

Significance:  In order to detect and identify pork adulteration in meat products and food is of concern to vegetarians and some religions such as Islam and Judaism. These results indicated that the indirect ELISA can be a useful tool for the rapid screening and quantification of pork fat adulteration in meat products.