Comparison of two assay methods for patterns of adherence to HEp-2 cells of Escherichia coli from patients with diarrhea

Abstract

To determine whether methodological differences in the HEp-2 adherence assay could explain conflicting results of field studies, 244 strains of Escherichia coli from Mexican children with diarrhea were tested for patterns of adherence by the method used at the Center for Vaccine Development, University of Maryland (CVD), and at the Center for Infectious Diseases, University of Texas Medical School and School of Public Health (UTH). The CVD assay differentiated three phenotypes of adherent E. coli, including localized, diffuse, or aggregative adherence (LA, DA, or AA, respectively). There was agreement on pattern of adherence in 241 of the 244 strains (98.8%) tested by the CVD method in both Baltimore and Houston, and AA+ was the most common phenotype (28.5% of isolates). Among these isolates, the UTH assay detected only two adherent phenotypes (LA and DA), since it did not distinguish the AA pattern. The LA+ strains detected by each assay were compared for positivity with the enteropathogenic E. coli adherence factor (EAF) gene probe. Of the 16 strains LA+ by the CVD method, 100% were EAF+; in contrast, only 11 of 22 strains LA+ by the UTH method were EAF+ (P = 0.00074). These results help explain why in pediatric field studies in Mexico where isolates were tested by the UTH method (J. J. Mathewson, R. A. Oberhelman, H. L. Dupont, F. J. de la Cabada, and E. V. Garibay, J. Clin. Microbiol. 25:1917-1919, 1987) LA+ strains often did not belong to enteropathogenic E. coli O serogroups and why the AA pattern was not observed; the opposite was found in studies of pediatric diarrhea in Chile in which the CVD assay was used (M. M. Levine, V. Prado, R. M. Robins-Browne, H. Lior, J. B. Kaper, S. Moseley, K. Gicquelais, J. P. Nataro, P. Vial, and B. Tall, J. Infect. Dis. 158:224-228, 1988). Since it appears that both assays identify E. coli strains associated with diarrheal illness, the genetic relationships among these strains should be examined in future studies.