Fluorescence based rRNA sensor systems for detection of whole cells of Saccharomonospora spp. and Thermoactinomyces spp |
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| Authors: | Neef Alexander Schäfer Rita Beimfohr Claudia Kämpfer Peter |
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| Institution: | 1. Institut für Angewandte Mikrobiologie, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 26-32, D-35392 Giessen, FRG;2. vermicon AG, Emmy-Noether-Str. 2, D-80992 Munich, FRG;1. Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, United States;2. Department of Immunology and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, United States;3. Worm Institute for Research Medicine (WIRM), The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, United States;1. Martin-Luther Universität Halle-Wittenberg, Bereich Organische Chemie, Kurt-Mothes-Str. 2, D-06120 Halle (Saale), Germany;2. Chiroblock GmbH, Andresenstr. 1a, D-06766 Bitterfeld-Wolfen, Germany;1. College of Pharmacy, Sookmyung Women’s University, Seoul 140-742, Republic of Korea;2. College of Pharmacy, Yeungnam University, Gyeongsan 712-749, Republic of Korea;3. College of Pharmacy, Graduate School of Pharmaceutical Sciences, and Global Top 5 Research Program, Ewha Womans University, Seoul 120-750, Republic of Korea;1. Groupe de Spectrometrie Moléculaire et Atmospherique, UMR CNRS 7331, University of Reims Champagne-Ardenne, F-51687 Reims Cedex 2, France;2. Department of Physics, University of Central Florida, Orlando, FL 32816, USA |
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| Abstract: | Airborne thermophilic actinomycetes (TPAs) are a growing hygienic challenge in different occupational situations e.g. large scale composting. This study describes first results of a new approach for highly specific and rapid detection of organisms of this group using fluorescently labelled oligonucleotide probes as sensors for whole cells. Three genus-specific 16S rRNA-targeted probes, two for Saccharomonospora spp. and one for Thermoactinomyces spp. were developed and evaluated in a fluorescence in situ hybridisation (FISH) format with agar-grown whole cells. For optimal sensitivity and specificity of FISH, conditions for cell wall permeabilisation and hybridisation stringency were evaluated independently for both genera. Performing specified pretreatment protocols, all three probes yielded strong fluorescence signals. However, the relative fraction of detectable cells or spores clearly depended on the single bacterial species. The probes can serve as cell sensors for direct detection of TPAs in natural samples. |
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