Use of an immunoperoxidase method for identification of Bacteroides fragilis

Abstract

An indirect immunoperoxidase (IP) slide test was evaluated for the laboratory identification of Bacteroides fragilis. Antigen-antibody complexes were detected with goat anti-rabbit immunoglobulin G-peroxidase conjugate with 3-amino-9-ethyl-carbazole as the peroxidase substrate. Ninety-one percent of 44 B. fragilis strains tested were IP positive (3+ to 4+ reactions) with greater than or equal to 1:160 dilutions of rabbit antiserum produced against whole cells of B. fragilis ATCC 23745. The antiserum was species specific. No cross-reactions were observed with 35 Bacteroides strains of other species or with a variety of facultative or aerobic gram-negative bacilli. Four B. fragilis strains were IP negative. One of these (VPI 2393) was the deoxyribonucleic acid (DNA) homology group II reference strain. The other three were clinical isolates. IP-negative and representative IP-positive strains were tested for DNA homology with the type strains for DNA homology groups I and II (VPI 2553 and VPI 2393). Two of the three clinical isolates were classified as DNA homology group II, and the remaining strain was classified as a group I. Capsular material known to be unique to B. fragilis was common to both DNA homology groups as indicated by reactions with purified anticapsular antiserum. The IP technique provides a suitable alternative to fluorescent microscopy for the rapid immunological identification of B. fragilis.