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PMID: 54339 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Quantitative assay of diphtherial toxin and of immunologically cross-reacting proteins by reversed passive hemagglutination.

Infection and immunity ·Vol. 12 ·No. 6 ·1975-12-00 ·Pages 1392-400

Holmes RK, Perlow RB

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.

MeSH Terms
Bacterial Proteins/analysis,immunology Corynebacterium diphtheriae/immunology Cross Reactions Diphtheria Antitoxin Diphtheria Toxin/analysis Epitopes Hemagglutination Hemagglutination Inhibition Tests Hemagglutination Tests Immunodiffusion
Chemicals
Bacterial Proteins Diphtheria Antitoxin Diphtheria Toxin Epitopes
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Holmes R K
Perlow R B
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33 references, click to expand
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Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/iai.12.6.1392-1400.1975
Published
1975-12-00
Pages
1392-400
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC415447
Subset
IM
External Links
PubMed source DOI: 10.1128/iai.12.6.1392-1400.1975 PMC Full Text View journal details
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