Screening of pig intestines for K88 non-adhesive phenotype by enzyme immunoassay

Abstract

An adhesion test for binding of porcine brush border membranes to Escherichia coli cells that possess the K88 antigen (K88 +) has been developed using enzyme immunoassay procedures. K88 pilus protein or K88 + E. coli cells were immobilized in the wells of polystyrene microtitre plates. These plates were incubated in the presence of material obtained by scraping the villous surface of pig small intestines. Adhesion of membrane material to immobilized K88 was detected by adding rabbit anti-brush border IgG followed by urease-labelled sheep anti-rabbit IgG conjugate. Action of bound enzyme on urea/bromo-cresol purple substrate solution (pH 4.8) produced an intense colour change from yellow to purple, enabling the test to be read visually. This test enables simple, rapid testing of large numbers of intestial samples and gives results that agree well with the more cumbersome microscopic adhesion test for adhesion of K88 + E. coli to purified brush border membranes.