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. 1995 Jan;61(1):13–20. doi: 10.1128/aem.61.1.13-20.1995

Development and application of a monoclonal antibody against Thiothrix spp.

R L Brigmon 1, G Bitton 1, S G Zam 1, B O'Brien 1
PMCID: PMC167257  PMID: 7887596

Abstract

Historically, methods used to identify Thiothrix spp. in environmental samples have been inadequate because isolation and identification procedures are time-consuming and often fail to separate Thiothrix spp. from other filamentous microorganisms. We described a monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) procedure which was used to identify Thiothrix spp. in wastewater, artesian springs, groundwater, and underwater subterranean samples. The ELISA utilized monoclonal antibody T3511 to a species-specific carbohydrate epitope of Thiothrix spp. No cross-reactions were observed among non-Thiothrix strains consisting of 12 species and nine genera. In field trials, the ELISA identified 100% of 20 biochemically and cytologically confirmed Thiothrix spp.-containing samples with no false positives. Indirect immunofluorescent microscopy utilizing T3511 was effective for wastewater samples but not for those from natural spring water because of background fluorescence in the latter. In addition, electron micrographs of Thiothrix spp. labeled with T3511-biotin-anti-mouse antibody-gold showed that epitope T3511 was intracellular both in laboratory strains and environmental isolates. The minimum level of detection of the ELISA was 0.10 microgram/ml.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amann R. I., Krumholz L., Stahl D. A. Fluorescent-oligonucleotide probing of whole cells for determinative, phylogenetic, and environmental studies in microbiology. J Bacteriol. 1990 Feb;172(2):762–770. doi: 10.1128/jb.172.2.762-770.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brigmon R. L., Zam S. G., Bitton G., Farrah S. R. Detection of Salmonella enteritidis in environmental samples by monoclonal antibody-based ELISA. J Immunol Methods. 1992 Jul 31;152(1):135–142. doi: 10.1016/0022-1759(92)90096-c. [DOI] [PubMed] [Google Scholar]
  3. Farquhar G. J., Boyle W. C. Control of Thiothrix in activated sludge. J Water Pollut Control Fed. 1972 Jan;44(1):14–24. [PubMed] [Google Scholar]
  4. Farquhar G. J., Boyle W. C. Occurrence of filamentous microorganisms in activated sludge. J Water Pollut Control Fed. 1971 May;43(5):779–798. [PubMed] [Google Scholar]
  5. HAROLD R., STANIER R. Y. The genera Leucothrix and Thiothrix. Bacteriol Rev. 1955 Jun;19(2):49–64. doi: 10.1128/br.19.2.49-64.1955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Johnson G. D., Davidson R. S., McNamee K. C., Russell G., Goodwin D., Holborow E. J. Fading of immunofluorescence during microscopy: a study of the phenomenon and its remedy. J Immunol Methods. 1982 Dec 17;55(2):231–242. doi: 10.1016/0022-1759(82)90035-7. [DOI] [PubMed] [Google Scholar]
  7. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  8. Larkin J. M., Strohl W. R. Beggiatoa, Thiothrix, and Thioploca. Annu Rev Microbiol. 1983;37:341–367. doi: 10.1146/annurev.mi.37.100183.002013. [DOI] [PubMed] [Google Scholar]
  9. McGlannan M. F., Makemson J. C. HCO(3) Fixation by Naturally Occurring Tufts and Pure Cultures of Thiothrix nivea. Appl Environ Microbiol. 1990 Mar;56(3):730–738. doi: 10.1128/aem.56.3.730-738.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Shuttleworth K. L., Unz R. F. Sorption of heavy metals to the filamentous bacterium thiothrix strain A1. Appl Environ Microbiol. 1993 May;59(5):1274–1282. doi: 10.1128/aem.59.5.1274-1282.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Steward M. W., Lew A. M. The importance of antibody affinity in the performance of immunoassays for antibody. J Immunol Methods. 1985 Apr 22;78(2):173–190. doi: 10.1016/0022-1759(85)90074-2. [DOI] [PubMed] [Google Scholar]
  12. Williams T. M., Unz R. F. Filamentous sulfur bacteria of activated sludge: characterization of Thiothrix, Beggiatoa, and Eikelboom type 021N strains. Appl Environ Microbiol. 1985 Apr;49(4):887–898. doi: 10.1128/aem.49.4.887-898.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. van Veen W. L. Bacteriology of activated sludge, in particular the filamentous bacteria. Antonie Van Leeuwenhoek. 1973;39(2):189–205. doi: 10.1007/BF02578852. [DOI] [PubMed] [Google Scholar]

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