Production, characterization, and application of antibodies against heat-labile type-I toxin for detection of enterotoxigenic Escherichia coli.
Strains of enterotoxigenic Escherichia coli (ETEC) are responsible for significant rates of morbidity and mortality among children, particularly in developing countries. The majority of clinical and public health laboratories are capable of isolating and identifying Salmonella, Shigella, Campylobacter, and Escherichia coli O157:H7 from stool samples, but ETEC cannot be identified by routine methods. The method most often used to identify ETEC is polymerase chain reaction for heat-stable and heat-labile enterotoxin genes, and subsequent serotyping, but most clinical and public health laboratories do not have the capacity or resources to perform these tests. In this study, polyclonal rabbit and monoclonal mouse IgG2b antibodies against ETEC heat-labile toxin-I (LT) were characterized and the potential applicability of a capture assay was analyzed. IgG-enriched fractions from rabbit polyclonal and the IgG2b monoclonal antibodies recognized LT in a conformational shape and they were excellent tools for detection of LT-producing strains. These findings indicate that the capture immunoassay could be used as a diagnostic assay of ETEC LT-producing strains in routine diagnosis and in epidemiological studies of diarrhea in developing countries as enzyme linked immunosorbent assay techniques remain as effective and economical choice for the detection of specific pathogen antigens in cultures.
Menezes
Caroline A
CA
Laboratório de Bacteriologia, Instituto Butantan, 05503-900 São Paulo, SP, Brasil.
Imamura
Sergio Y
SY
Trabulsi
Luiz R
LR
Fernandes-Filho
Antônio
A
Martinez
Marina B
MB
Guth
Beatriz E C
BE
Girão
Dennys M
DM
Piazza
Roxane M F
RM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D001427:Bacterial Toxins / Q000276:immunology*; D002648:Child; D004768:Enterotoxins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology*; D029968:Escherichia coli Proteins; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000096:biosynthesis*; D051379:Mice; D016133:Polymerase Chain Reaction; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012703:Serotyping
D000911:Antibodies, Monoclonal; D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D007074:Immunoglobulin G; C034151:heat stable toxin (E coli)