Solid-phase radioimmunoassay method for determination of Escherichia coli enterotoxin

Ceska, M.; Grossmüller, F.; Effenberger, Friedrich

doi:10.1128/iai.19.2.347-352.1978

Cited by 10 publications

(4 citation statements)

References 15 publications

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“…A good method should be simple, accurate, reproducible, cheap, and sensitive. In general, immunological methods are better than others, and of the various immunological methods so far reported (3,5,6,9,21,27,31), ELISA is the most widely used, because it is the most sensitive, accurate, and reproducible. The sensitivities of the ELISA's reported previously, however, varied significantly.…”

Section: Discussionmentioning

confidence: 99%

Development of a Highly Sensitive Bead‐ELISA to Detect Bacterial Protein Toxins

Oku

Uesaka

Hirayama

et al. 1988

Microbiology and Immunology

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A highly sensitive sandwich enzyme-linked immunosorbent assay to detect bacterial toxins was developed. Fab' of anti-toxin IgG was conjugated with horseradish peroxidase by the maleimide method and tetramethylbenzidine was used as substrate. As the solid phase, a 6.5 mm diameter polystyrene bead was used and this was coated with the anti-toxin IgG. The entire assay could be completed within 3.5 hr. The sensitivity of this bead-ELISA was found to be quite high with various bacterial toxins : less than 20 pg/ml for thermostable direct hemolysin of Vibrio parahaemolyticus, less than 60 pg/ml for Shiga toxin, less than 20 pg/ml for VT2 (Shiga-like toxin II) of Escherichia coli, less than 200 pg/ml for heat-labile enterotoxin of E. coli, and less than 6 pg/ml for cholera enterotoxin.

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Section: Discussionmentioning

confidence: 99%

Development of a Highly Sensitive Bead‐ELISA to Detect Bacterial Protein Toxins

Oku

Uesaka

Hirayama

et al. 1988

Microbiology and Immunology

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“…As LT was then not yet available in purified form, pure cholera toxin was used for the cross reaction of antiserum to LT. Solid phase immunoassay was also used by Ceska et al (1978).…”

Section: (I) In Vivo Assay In Ligated Intestinal Loopsmentioning

confidence: 99%

Enterotoxins from gram-negative bacteria relevant for veterinary medicine

Rašková

Raska

1980

Vet Res Commun

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“…The first generation of such tests employed clinically consisted of tissue culture assays for LT and the suckling mouse assay for ST in broth cultures of isolates (36). Other techniques were subsequently introduced for assaying LT in broth cultures; some, such as passive immune hemolysis (12) and radioimmunoassay (5,17), proved too complex for adoption in general use. More recently, simple, sensitive enzyme-linked immunosorbent assays (ELISAs) for LT, based on the use of ganglioside GM1 as the solid phase and hyperimmune antiserum to LT as the second antibody (GM1-LT), have been developed (3,9,19,35,42,51,52).…”

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confidence: 99%

Enzyme-linked immunosorbent assay for Escherichia coli heat-stable enterotoxin

Klipstein¹,

Engert²,

Houghten³

et al. 1984

J Clin Microbiol

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The sensitivity of an enzyme-linked immunosorbent assay (ELISA) to detect pure native Escherichia coli heat-stable toxin (ST) and to identify ST-producing strains among clinical isolates was determined. Two synthetically produced ST preparations were used to raise hyperimmune antisera in rabbits and goats: ST(S), which has the same antigenicity as native ST; and ST(C), which is 15-fold more immunogenic. These antisera were used in the double-sandwich technique as either crude double-species antisera or pure singlespecies antibody. The sensitivity of the assay was increased by using either a purer antibody preparation or the antiserum to the more potent immunogen; the assay in which pure antibody to ST(C) was used was 2,857-fold more sensitive in detecting ST than the assay in which crude antiserum to ST(S) was used. The minimum amount of ST detectable by the ST(C) ELISA was 140 pg/ml, which was an amount 285-fold smaller than that detectable by the suckling mouse assay. Among 50 human E. coli isolates examined by both the ST(C) ELISA and an ELISA for heat-labile toxin (LT), which had a sensitivity of 290 pg/ml for LT, the respective toxins were consistently identified in broth cultures of 10 LT' and ST-, 15 LT' and ST+, and 10 LT-and ST' strains, and there were no false-positive responses. The ST(C) ELISA also detected ST in all of seven ST-producing E. coli strains tested of human origin, which had been shown elsewhere by DNA hybridization probes to have ST-coding genes of either human or porcine origin, and in all of three ST-producing E. coli strains tested of porcine origin. These results indicate that the sensitivity of the ST(C) ELISA is the same as that of previously described LT ELISAs. The concomitant use of both ST and LT ELISAs provides a rapid, simple, and sensitive method for identifying among clinical isolates enterotoxigenic strains of E. coli which produce either toxin.

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Solid-phase radioimmunoassay method for determination of Escherichia coli enterotoxin

Abstract: Polyacrylamide gel electrophoresis of radioiodinated E. coli enterotoxin. The total concentration of polyacrylamide in the separation gel was 347 on August 5, 2020 by guest http://iai.asm.org/ Downloaded from

Cited by 10 publications

References 15 publications

Development of a Highly Sensitive Bead‐ELISA to Detect Bacterial Protein Toxins

Development of a Highly Sensitive Bead‐ELISA to Detect Bacterial Protein Toxins

Enterotoxins from gram-negative bacteria relevant for veterinary medicine

Enzyme-linked immunosorbent assay for Escherichia coli heat-stable enterotoxin

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