Molecular Genetic Methods for Detection and Identification of Viable but Nonculturable Microorganisms
The problems associated with culture-based methods for detection and identification of microorganisms in clinical and environmental samples have motivated the development of alternative methods which do not require cultivation of the target organisms. Molecular genetic methods, which specifically target microbial nucleic acids, have become important tools for the identification of both cultured and uncultured microorganisms. The sensitivity of these methods approaches that of culture-based methods and it is arguable that the specificity of these methods generally exceeds that of culture-based methods. Indeed, molecular genetic methods have become a powerful set of tools used by many investigators to detect and identify culturable microorganisms in their viable but nonculturable state, as well as organisms which have yet to be brought into culture.
KeywordsMethane Sludge Tuberculosis Electrophoresis Bacillus
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- 8.Bogert, A. P., and I. T. Knight. 1995. Detection of enterotoxigenic E. coli in ground water using DNA hybridization and PCR, abstr. N-76, p. 345. In Abstracts of the 95th General Meeting of the American Society for Microbiology 1995. American Society for Microbiology, Washington, D.C.Google Scholar
- 15.Hicks, R. E., R. I. Amann, and D. A. Stahl. 1991. Dual staining of natural bacterioplankton with 4′,6-diamidino-2-phenylindole and fluorescent oligonucleotide probes targeting kingdom-level 16S rRNA sequences. Appl. Environ. Microbiol. 58:2158–2163.Google Scholar
- 17.Islam, M. S., M. K. Hasan, M. A. Miah, G. C. Sur, A. Felsenstein, M. Venkatesan, R. B. Sack, and M. J. Albert. 1993. Use of the polymerase chain reaction and fluorescent-antibody methods for detecting viable but nonculturable Shigella dysenteriae type 1 in laboratory microcosms. Appl. Environ. Microbiol. 59:536–540.PubMedGoogle Scholar
- 21.Knight, I. T., J. DiRuggiero, and R. R. Colwell. 1991. Direct detection of enteropathogenic bacteria in estuarine water using nucleic acid probes. Water Sci. Technol. 24:261–266.Google Scholar
- 22.Knight, I. T., W. E. Holben, J. M. Tiedje, and R. R. Colwell. 1991. Nucleic acid hybridization techniques for detection, identification and enumeration of microorganisms in the environment, p. 65–91. In M. Levin, R. J. Seidler, and M. Rogul (ed.), Microbial Ecology. Principles, Methods and Application to Environmental Biotechnology. McGraw-Hill, Inc., New York, N.Y.Google Scholar
- 27.Manz, W., U. Szewzyk, P. Ericsson, R. Amann, K.-H. Schleifer, and T.-A. Stenström. 1993. In situ identification of bacteria in drinking water and adjoining biofilms by hybridization with 16S and 23S rRNA-directed fluorescent oligonucleotide probes. Appl. Environ. Microbiol. 59:2293–2298.PubMedGoogle Scholar
- 34.Nuovo, G. J. 1994. In situ detection of PCR-amplified DNA and cDNA: a review. J. Histotechnol. 17:235–246.Google Scholar
- 55.Wagner, M., R. Erhart, W. Manz, R. Amann, H. Lemmer, D. Wedi, and K.-H. Schleifer. 1994. Development of an rRNA-targeted oligonucleotide probe specific for the genus Acinetobacter and its application for in situ monitoring in activated sludge. Appl. Environ. Microbiol. 60:792–800.PubMedGoogle Scholar