Abstract
To detect low levels of microorganisms in environmental samples by using polymerase chain reaction (PCR)-gene probe detection, samples were concentrated by filtration. Fluoropore (Millipore Corp.) filters were compatible with PCR DNA amplification, whereas various other filters including nitrocellulose and cellulose acetate filters inhibited PCR amplification. By concentrating cells on Fluoropore filters and releasing the DNA by freeze-thaw cycling, PCR DNA amplification could be performed without removing the filter. Concentration with Fluoropore FHLP and FGLP filters permitted the detection of single cells of microorganisms in 100-ml samples by PCR-gene probes.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bej A. K., DiCesare J. L., Haff L., Atlas R. M. Detection of Escherichia coli and Shigella spp. in water by using the polymerase chain reaction and gene probes for uid. Appl Environ Microbiol. 1991 Apr;57(4):1013–1017. doi: 10.1128/aem.57.4.1013-1017.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bej A. K., Mahbubani M. H., Miller R., DiCesare J. L., Haff L., Atlas R. M. Multiplex PCR amplification and immobilized capture probes for detection of bacterial pathogens and indicators in water. Mol Cell Probes. 1990 Oct;4(5):353–365. doi: 10.1016/0890-8508(90)90026-v. [DOI] [PubMed] [Google Scholar]
- 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]
- Bopp C. A., Sumner J. W., Morris G. K., Wells J. G. Isolation of Legionella spp. from environmental water samples by low-pH treatment and use of a selective medium. J Clin Microbiol. 1981 Apr;13(4):714–719. doi: 10.1128/jcm.13.4.714-719.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dondero T. J., Jr, Rendtorff R. C., Mallison G. F., Weeks R. M., Levy J. S., Wong E. W., Schaffner W. An outbreak of Legionnaires' disease associated with a contaminated air-conditioning cooling tower. N Engl J Med. 1980 Feb 14;302(7):365–370. doi: 10.1056/NEJM198002143020703. [DOI] [PubMed] [Google Scholar]
- Fitzgeorge R. B., Dennis P. J. Isolation of Legionella pneumophila from water supplies: comparison of methods based on the guinea-pig and culture media. J Hyg (Lond) 1983 Oct;91(2):179–187. doi: 10.1017/s0022172400060186. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grossman N., Ron E. Z. Membrane-bound DNA from Escherichia coli: extraction by freeze-thaw-lysozyme. FEBS Lett. 1975 Jul 1;54(3):327–329. doi: 10.1016/0014-5793(75)80932-x. [DOI] [PubMed] [Google Scholar]
- Kilian M., Bülow P. Rapid diagnosis of Enterobacteriaceae. I. Detection of bacterial glycosidases. Acta Pathol Microbiol Scand B. 1976 Oct;84B(5):245–251. doi: 10.1111/j.1699-0463.1976.tb01933.x. [DOI] [PubMed] [Google Scholar]
- Lewis C. M., Mak J. L. Comparison of membrane filtration and Autoanalysis Colilert presence-absence techniques for analysis of total coliforms and Escherichia coli in drinking water samples. Appl Environ Microbiol. 1989 Dec;55(12):3091–3094. doi: 10.1128/aem.55.12.3091-3094.1989. [DOI] [PMC free article] [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]
- Riggs J. L., Dupuis K. W., Nakamura K., Spath D. P. Detection of Giardia lamblia by immunofluorescence. Appl Environ Microbiol. 1983 Feb;45(2):698–700. doi: 10.1128/aem.45.2.698-700.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ron E. Z., Kohler R. E., Davis B. D. Polysomes extracted from Escherichia coli by freeze-thaw-lysozyme lysis. Science. 1966 Sep 2;153(3740):1119–1120. doi: 10.1126/science.153.3740.1119. [DOI] [PubMed] [Google Scholar]
- Rose J. B., Landeen L. K., Riley K. R., Gerba C. P. Evaluation of immunofluorescence techniques for detection of Cryptosporidium oocysts and Giardia cysts from environmental samples. Appl Environ Microbiol. 1989 Dec;55(12):3189–3196. doi: 10.1128/aem.55.12.3189-3196.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
- Sauch J. F. Use of immunofluorescence and phase-contrast microscopy for detection and identification of Giardia cysts in water samples. Appl Environ Microbiol. 1985 Dec;50(6):1434–1438. doi: 10.1128/aem.50.6.1434-1438.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]