Development and Validation of a Rapid Test Kit for Detection of Pork Meat Residues

Introduction: A recent investigation into meat adulteration revealed that many foods advertised as containing beef were found to contain undeclared pork residues. Whereas consumption of pork is not harmful, certain ethnic and religious practices forbid the consumption of pork meat. Moreover, undeclared pork meat can present a health risk if consumed in an undercooked state. In an effort to safeguard the public from inadvertent exposure to pork meat, we developed a rapid and highly specific test kit capable of detecting trace levels of pork meat residues.

Purpose: To develop and validate a highly specific detection kit that can rapidly identify pork meat residues down to 0.01% contamination in xenogenic meat sources.

Methods: Polyclonal antibodies against pork serum albumin (PSA) were raised in goats and purified on a PSA-affinity column. The purified antibodies were then used to develop a flow immunochromatographic assay configured with a procedural control line, a test line, and an overload line.  A sample extraction buffer and a running buffer were developed for optimal performance. The ensuing kit and test method were validated for specificity and dynamic range. Method concordance was assessed using a commercial ELISA (ELISA-TEK) kit and results were confirmed using a validated meat authentication PCR-based method (IEH).

Results: The MEI/IEH pork meat lateral flow test demonstrated a range of detection of 0.01% to 100% spiked pork meat (raw or cooked meat into beef meat). Specificity analysis revealed no cross-reactivity with serum albumins or meats derived from chicken, turkey, horse, beef, lamb, and goat. The assay was more sensitive than the commercial ELISA kit and required considerably less time than the PCR and ELISA methods.

Significance: The development of a highly specific test method capable of detecting trace amounts of horse meat in 30 min should aid the Food Safety Authorities in their continued efforts to monitor for pork meat adulteration in the food chain. Furthermore, the facile operation of this kit should enable religious parties the opportunity to independently perform meat authentication testing.