Quantitative Assay of Diphtherial Toxin and of Immunologically Cross-reacting Proteins by Reversed Passive Hemagglutination

Overview

Journal Infect Immun

Publisher American Society for Microbiology

Specialty Allergy & Immunology

Date 1975 Dec 1

PMID 54339

Citations 14

Authors

R K Holmes

R B Perlow

Affiliations

Soon will be listed here.

Abstract

A reversed passive hemagglutination (RPHA) assay for diptherial toxin has been developed. Antitoxic antibodies were isolated from commercially available equine diptherial antitoxin by immunoabsorption using highly purified diphtherial toxin covalently linked to Sepharose 4B. Formalinized, tanned sheep erythrocytes sensitized with the purified antitoxic antibodies are specifically agglutinated by diphtherial toxin but are not agglutinated by extracellular antigens of Corynebacterium diptheriae that are unrelated to toxin. The RPHA assay described can detect less than 20 pg of diphtherial toxin and is comparable in sensitivity to intracutaneous tests for toxin. The RPHA assay was shown to be at least 1,000 times more sensitive than quantitative immunological assays for diptherial toxin performed by single radial immunodiffusion or by one-dimensional double diffusion in agar gels. Fragment A prepared from purified diphtherial toxin and nontoxic mutant proteins that cross-react immunologically with toxin can be assayed directly by RPHA, but the sensitivity of the assay for these proteins is less than for native diphtherial toxin. Inhibition of RPHA was also shown to be a sensitive quantitative method for measuring diptherial antitoxin in vitro.

Citing Articles

Challenges of Diphtheria Toxin Detection.

Prygiel M, Mosiej E, Polak M, Krysztopa-Grzybowska K, Wdowiak K, Forminska K Toxins (Basel). 2024; 16(6).

PMID: 38922140 PMC: 11209151. DOI: 10.3390/toxins16060245.

Inhibition of endoplasmic reticulum-associated degradation in CHO cells resistant to cholera toxin, Pseudomonas aeruginosa exotoxin A, and ricin.

Teter K, Holmes R Infect Immun. 2002; 70(11):6172-9.

PMID: 12379695 PMC: 130429. DOI: 10.1128/IAI.70.11.6172-6179.2002.

Reversed passive latex agglutination assay for detection of toxigenic Corynebacterium diphtheriae.

Toma C, Sisavath L, Iwanaga M J Clin Microbiol. 1997; 35(12):3147-9.

PMID: 9399510 PMC: 230138. DOI: 10.1128/jcm.35.12.3147-3149.1997.

Regulation of toxinogenesis in Corynebacterium diphtheriae. I. Mutations in bacteriophage beta that alter the effects of iron on toxin production.

Welkos S, Holmes R J Virol. 1981; 37(3):936-45.

PMID: 6785449 PMC: 171091. DOI: 10.1128/JVI.37.3.936-945.1981.

Regulation of toxinogenesis in Corynebacterium diphtheriae: mutations in the bacterial genome that alter the effects of iron on toxin production.

Cryz Jr S, Russell L, Holmes R J Bacteriol. 1983; 154(1):245-52.

PMID: 6403502 PMC: 217453. DOI: 10.1128/jb.154.1.245-252.1983.

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