Abstract
A method for toxigenicity testing of Corynebacterium diphtheriae in tissue cultures was developed. Results were obtained by comparing destruction of the monkey kidney or, preferably, rabbit kidney monolayer by 0.1 ml of the C. diphtheriae culture in Elek's broth containing 20% rabbit serum with the appearance after the addition of 0.2 ml of a mixture of the C. diphtheriae culture and diphtheria antitoxin. The mixture of C. diphtheriae broth culture and 10 antitoxin units per ml was incubated for 1 hr at room temperature before it was added to the tissue cultures which were then incubated as long as 5 days; most results, however, were read in 72 hr. Elek's broth medium was superior to heart infusion broth for toxin production by C. diphtheriae. Addition of 20% rabbit serum improved toxin production in either broth. Numerous toxigenic and atoxigenic C. diphtheriae cultures were tested for toxigenicity in primary rabbit and monkey kidney tissue cultures. If properly controlled, this in vitro method appeared to have an advantage over the in vitro agar gel method; its results were comparable with the rabbit intradermal test. With the wider use of tissue cultures in most laboratories, we believe that the tissue culture method for toxigenicity would be more economical and easier to perform than the animal intradermal method.
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- ANDRE J., AUDEBAUD G., CHAMBON L. [Diagnosis of diphtheria on tissue culture]. Ann Inst Pasteur (Paris) 1960 Aug;99:179–187. [PubMed] [Google Scholar]
- ELEK S. D. The plate virulence test for diphtheria. J Clin Pathol. 1949 Nov;2(4):250–258. doi: 10.1136/jcp.2.4.250. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LENNOX E. S., KAPLAN A. S. Action of diphtheria toxin on cells cultivated in vitro. Proc Soc Exp Biol Med. 1957 Aug-Sep;95(4):700–702. doi: 10.3181/00379727-95-23335. [DOI] [PubMed] [Google Scholar]
- ORMAY L., UJHELYI K. Titration of diphtheria toxin, antitoxin and toxoid in tissue culture. Acta Microbiol Acad Sci Hung. 1961;8:389–396. [PubMed] [Google Scholar]
- STRAUSS N., HENDEE E. D. The effect of diphtheria toxin on the metabolism of HeLa cells. J Exp Med. 1959 Feb 1;109(2):145–163. doi: 10.1084/jem.109.2.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schubert J. H., Wiggins G. L., Taylor G. C. Tissue culture method for the titration of diphtheria antitoxin in human sera. Health Lab Sci. 1967 Jul;4(3):181–188. [PubMed] [Google Scholar]
- Stănică E., Stoian C., Maximesco P., Stoian I., Calalb G. Contribution au titrage de la toxine-antitoxine diphtérique sur cultures de tissus. Arch Roum Pathol Exp Microbiol. 1964 Dec;23(4):1017–1024. [PubMed] [Google Scholar]