Speciation and Characterization of Beef by ORBIT, ELISA, and PAGE

One important aspect of the National Laboratories of Foods and Drugs (NLFD) analysis concerns the accurate identification of the bovine tissue used in consumable meat to assure that there is no adulteration or fraudulent substitution. Two immunological screening methods and an electrophoretic method, overnight rapid beef identification test(ORBIT), enzymelinked immuno-sorbent assay(ELISA) as well as polyacrylamide gel electrophoresis(PAGE) for beef detection, are applied to speciate meat species and characterization. Meat extracts including beef, buffalo, sheep, goat, kangaroo, donkey, horse, pig, Formosan spotted deer(梅花鹿) and Swinhoe's deer(水鹿) were used in this experiment. The subsidiary bands of myoglobin for species specific were identified and characterized by one, two dimensional and isoelectric focusing PAGE. The ORBIT is an example of a highly specific antigen-antibody immunodiffusion reaction, which could form a clearly visible white precipitin band if the concentration of beef is up to 10% or more. The ELISA is another highly specific immunological method, which could develop a blue color as a positive result if the sample's concentration is more than 1%. It is obviously that PAGE is the unique method for speciation in all species of meat including denatured one, but the ORBIT and ELISA do suitable for detecting the flesh in compliance with the proper antibody. Through the experiment described in this article the best strategy for identifying meat adulteration and the concentrationreaction time relationship curve will also be discussed.