This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Unson, M. D.
Right arrow Articles by Fahey, R. C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Unson, M. D.
Right arrow Articles by Fahey, R. C.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, July 1999, p. 2153-2157, Vol. 37, No. 7
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Improved Methods for Immunoassay of Mycothiol

Mia D. Unson,1 Gerald L. Newton,1 Karen F. Arnold,2 Charles E. Davis,2 and Robert C. Fahey1,*

Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, California 92093,1 and Department of Pathology, School of Medicine, University of California, San Diego, San Diego, California 921032

Received 29 December 1998/Returned for modification 27 February 1999/Accepted 26 March 1999

Improved enzyme-linked immunosorbent assay (ELISA) methods have been developed for the determination of femtomole amounts of mycothiol (MSH), the main low-molecular-weight thiol in mycobacteria. The immunoassays utilize an affinity-purified rabbit polyclonal antibody that is highly specific for the pseudodisaccharide moiety of MSH. MSH was first biotinylated by the thiol-specific reagent 3-(N-maleimidopropionyl)biocytin. The MSH-biotin adduct was then captured with immobilized avidin and detected with anti-MSH antibody (biotin-capture ELISA) or was captured with immobilized anti-MSH antibody and detected with alkaline phosphatase-labelled avidin (MSH-capture ELISA). The MSH-capture ELISA was the most sensitive method, measuring as little as 0.3 fmol of MSH. Methods for biotinylating MSH directly from Mycobacterium spp. are described. The MSH-capture ELISA was tested for the detection of M. avium seeded in human urine or cerebrospinal fluid samples and for screening mutant M. smegmatis strains to detect MSH production.

* Corresponding author. Mailing address: Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA 92093-0506. Phone: (619) 534-2163. Fax: (619) 534-4864. E-mail: rcfahey{at}ucsd.edu.

Journal of Clinical Microbiology, July 1999, p. 2153-2157, Vol. 37, No. 7
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Rawat, M., Newton, G. L., Ko, M., Martinez, G. J., Fahey, R. C., Av-Gay, Y. (2002). Mycothiol-Deficient Mycobacterium smegmatis Mutants Are Hypersensitive to Alkylating Agents, Free Radicals, and Antibiotics. Antimicrob. Agents Chemother. 46: 3348-3355 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»