Clinical Professor, Cardiology University of Florida Gainesville, Florida, United States
Abstract:
Background:
PHA is a lectin found in kidney beans and other legumes that is toxic if improperly processed during food manufacturing. PHA consists of two closely related subunits, PHA-E and PHA-L. The E and L subunits are responsible for erythroagglutination (red blood cell aggregation) and leucoagglutination (white blood cell aggregation) respectively. Increased levels of PHA-E originating from legumes in grain-free (GF) dog foods could theoretically cause DCM from red cell agglutination causing cardiac muscle strain. Legume-rich, GF dog foods are in the process of being evaluated for lectins.
Objective:
We sought to develop an ELISA to measure levels of Phytohemagglutinin (PHA) in legumes that are used in GF dog foods.
Method:
For the development of the ELISA, PHA-E analyte was coated onto 96 well microtiter plates. Antibody against the analyte was produced through the immunization of rabbits. Thirty-two legume samples were ground up and extracted in a 0.9% NaCl buffer to test for PHA-E by ELISA. The standard curve of the assay ranges from 1 to 100 ppm.
Results:
The 2 samples yielding the highest PHA-E levels were Red Kidney Bean at 35,204 ppm and Dark Kidney Bean at 14,480 ppm. Samples with medium PHA-E levels; Yellow Eyed Peas at 4,676 ppm and Navy Beans at 1290 ppm. Various Lentils tested had lower results ranging between 1 and 197 ppm.
Conclusion:
The ELISA developed in this study could be used to measure PHA-E in food which might contribute to our understanding of DCM associated with legume-rich GF dog foods.
