Abstract
Methods using polymerase chain reaction (PCR) and gene probes to detect viable Legionella pneumophila were investigated with cells exposed to biocide or elevated temperature. Exposure to hypochlorite caused viable nonculturable cells to form. Culturable and viable nonculturable cells showed positive PCR amplification, whereas nonviable cells did not. Viable cells were also specifically detected with mip mRNA as the target, reverse transcription (to form cDNA), and PCR amplification. After exposure to elevated temperature, only viable culturable cells were detected, which corresponded with positive PCR amplification.
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- Bej A. K., Steffan R. J., DiCesare J., Haff L., Atlas R. M. Detection of coliform bacteria in water by polymerase chain reaction and gene probes. Appl Environ Microbiol. 1990 Feb;56(2):307–314. doi: 10.1128/aem.56.2.307-314.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Belasco J. G., Higgins C. F. Mechanisms of mRNA decay in bacteria: a perspective. Gene. 1988 Dec 10;72(1-2):15–23. doi: 10.1016/0378-1119(88)90123-0. [DOI] [PubMed] [Google Scholar]
- England A. C., 3rd, Fraser D. W., Mallison G. F., Mackel D. C., Skaliy P., Gorman G. W. Failure of Legionella pneumophila sensitivities to predict culture results from disinfectant-treated air-conditioning cooling towers. Appl Environ Microbiol. 1982 Jan;43(1):240–244. doi: 10.1128/aem.43.1.240-244.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Engleberg N. C., Carter C., Weber D. R., Cianciotto N. P., Eisenstein B. I. DNA sequence of mip, a Legionella pneumophila gene associated with macrophage infectivity. Infect Immun. 1989 Apr;57(4):1263–1270. doi: 10.1128/iai.57.4.1263-1270.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Feeley J. C., Gibson R. J., Gorman G. W., Langford N. C., Rasheed J. K., Mackel D. C., Baine W. B. Charcoal-yeast extract agar: primary isolation medium for Legionella pneumophila. J Clin Microbiol. 1979 Oct;10(4):437–441. doi: 10.1128/jcm.10.4.437-441.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Higuchi R., von Beroldingen C. H., Sensabaugh G. F., Erlich H. A. DNA typing from single hairs. Nature. 1988 Apr 7;332(6164):543–546. doi: 10.1038/332543a0. [DOI] [PubMed] [Google Scholar]
- Impraim C. C., Saiki R. K., Erlich H. A., Teplitz R. L. Analysis of DNA extracted from formalin-fixed, paraffin-embedded tissues by enzymatic amplification and hybridization with sequence-specific oligonucleotides. Biochem Biophys Res Commun. 1987 Feb 13;142(3):710–716. doi: 10.1016/0006-291x(87)91472-0. [DOI] [PubMed] [Google Scholar]
- Mahbubani M. H., Bej A. K., Miller R. D., Atlas R. M., DiCesare J. L., Haff L. A. Detection of bacterial mRNA using polymerase chain reaction. Biotechniques. 1991 Jan;10(1):48–49. [PubMed] [Google Scholar]
- Mahbubani M. H., Bej A. K., Miller R., Haff L., DiCesare J., Atlas R. M. Detection of Legionella with polymerase chain reaction and gene probe methods. Mol Cell Probes. 1990 Jun;4(3):175–187. doi: 10.1016/0890-8508(90)90051-z. [DOI] [PubMed] [Google Scholar]
- Roszak D. B., Colwell R. R. Survival strategies of bacteria in the natural environment. Microbiol Rev. 1987 Sep;51(3):365–379. doi: 10.1128/mr.51.3.365-379.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tobin J. O., Beare J., Dunnill M. S., Fisher-Hoch S., French M., Mitchell R. G., Morris P. J., Muers M. F. Legionnaires' disease in a transplant unit: isolation of the causative agent from shower baths. Lancet. 1980 Jul 19;2(8186):118–121. doi: 10.1016/s0140-6736(80)90005-7. [DOI] [PubMed] [Google Scholar]