Application of a Multiplex PCR with Specific PCR Primers for the Detection of the Genus Paramecium and the Paramecium aurelia Complex
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RIS BIB ENDNOTEApplication of a Multiplex PCR with Specific PCR Primers for the Detection of the Genus Paramecium and the Paramecium aurelia Complex
Publication date: 15.05.2011
Acta Protozoologica, 2011, Volume 50, Issue 3, pp. 235-238
https://doi.org/10.4467/16890027AP.11.022.0022Authors
Application of a Multiplex PCR with Specific PCR Primers for the Detection of the Genus Paramecium and the Paramecium aurelia Complex
The representatives of the genus Paramecium are well-studied ciliates and can be used in water quality assessment and the determinations of saprobic levels. For these applications, a clear and unambiguous identification of ciliate assemblages is essential, which is typically based on morphological characters requiring a sound taxonomic knowledge and experience in species determination including microscopic identification of both living and stained specimens. Therefore, we developed and applied specific PCR primers for the detection of species belonging to the genus Paramecium and the Paramecium aurelia complex. These primers were successfully tested with different Paramecium species including representatives of the P. aurelia complex as well as closely related species like Frontonia sp. and Tetrahymena sp. in both experimental and environmental samples. These primers can be used in a simultaneous approach achieving fast and reliable results with regard to determination of ciliate community and water assessment.
Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. (1990) Basic local alignment search tool. J. Mol. Biol. 215: 403–410
Berger H., Foissner W., Kohmann F. (1997) Bestimmung und Ökologie der Mikrosaprobien nach DIN 38 410. G. Fischer, Stuttgart, Jena, Lübeck, Ulm
Catania F., Wurmser F., Potekhin A. A., Przybos E., Lynch M. (2009) Genetic diversity in the Paramecium aurelia species complex. Mol. Biol. Evol. 26: 421–431
Curds C. R., Cockburn A. (1970) Protozoa in biological sewagetreatment processes–II. Protozoa as indicators in the activatedsludge process. Water Res. 4: 237–249
Elwood H. J., Olsen G. J., Sogin M. L. (1985) The small-subunit ribosomal RNA gene sequences from the hypotrichous ciliates Oxytricha nova and Stylonychia pustulata. Mol. Biol. Evol. 2: 399–410
Epstein S., López-García P. (2008) “Missing” protists: a molecular prospective. Biodivers Conserv. 17: 261–276
Fokin S. I., Przybos E., Chivilev S. M., Beier C. L., Horn M., Skotarczak B., Wodecka B., Fujishima M. (2004) Morphological and molecular investigations of Paramecium schewiakoffi sp. nov. (Ciliophora, Oligohymenophorea) and current status of distribution and taxonomy of Paramecium spp. Eur. J. Protistol. 40: 225–243
Ghosh S., Debnath A., Sil A., De S., Chattopadhyay D. J., Das P. (2000) PCR detection of Giardia lamblia in stool: targeting intergenic spacer region of multicopy rRNA gene. Mol. Cell. Probes 14: 181–189
Haentzsch M., Schmidt S. L., Bernhard D., Ammermann D., Berendonk T. U., Schlegel M. (2006) A PCR-based method to distinguish the sibling species Stylonychia mytilus and Stylonychia lemnae (Ciliophora, Spirotrichea) using isocitrate dehydrogenase (IDH) gene sequences. J. Eukaryot. Microbiol. 53: 343– 347
Hide G., Hughes J. M., McNuff R. (2003) A rapid and simple method of detection of Blepharisma japonicum using PCR and immobilisation on FTA paper. BMC Ecol. 3: 7
Kalendar R., Lee D., Schulman A. H. (2009) FastPCR Software for PCR Primer and Probe Design and Repeat Search. Genes, Ge nomes and Genomics 3: 1–14
Monis P. T., Saint C. P. (2001) Development of a nested-PCR assay for the detection of Cryptosporidium parvum in finished water. Water Res. 35: 1641–1648
Petroni G., Rosati G., Vannini C., Modeo L., Dini F., Verni F. (2003) In situ identification by fluorescently labeled oligonucleotide probes of morphologically similar, closely related ciliate species. Microb. Ecol. 45: 156–162
Schmidt S. L., Bernhard D., Schlegel M., Fried J. (2006) Fluorescence in situ hybridization with specific oligonucleotide rRNA probes distinguish the sibling species Stylonychia lemnae and Stylonychia mytilus (Ciliophora, Spirotrichea). Protist 157: 21–30
Schmidt S. L., Treuner T., Schlegel M., Bernhard D. (2007) Multiplex PCR approach for species detection and differentiation within the genus Spirostomum (Ciliophora, Heterotrichea). Pro tist 158: 139–145
Thompson J. D., Gibson T. J., Plewniak F., Jeanmougin F., Higgins D. G. (1997) The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 25: 4876–4882
Udeh P., Veenstra J., Abraham A. J., John G. H. (2000) Quantitative polymerase chain (QPCR) reaction using the MIMIC approach to estimate Cryptosporidium parvum oocysts, an intestinal pathogen, in municipal water treatment sludge samples. Mol. Cell. Probes 14: 121–126
Wellinghausen N., Frost C., Marre R. (2001) Detection of Legionellae in hospital water samples by quantitative real-time LightCycler PCR. Appl. Environ. Microbiol. 67: 3985–3993
Information: Acta Protozoologica, 2011, Volume 50, Issue 3, pp. 235-238
Article type: Original article
Institute for Biology, Molecular Evolution and Animal Systematics, University of Leipzig, Leipzig, Germany
Institute for Hydrobiology, TU Dresden, Dresden, Germany
Institute for Hydrobiology, TU Dresden, Dresden, Germany
Department of Experimental Zoology, Institute of Systematics and Evolution of Animals, Polish Academy of Sciences, Kraków, Poland
Institute for Biology, Molecular Evolution and Animal Systematics, University of Leipzig, Leipzig, Germany
Published at: 15.05.2011
Article status: Open
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