title: Fluorescence based rRNA sensor systems for detection of whole cells of Saccharomonospora spp. and Thermoactinomyces spp.
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.
authors.affiliation: Institut für Angewandte Mikrobiologie, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 26-32, D-35392, Giessen, FRG. aln@gbf.de
authors.forename: Alexander
authors.initials: A
authors.lastname: Neef
authors.forename: Rita
authors.initials: R
authors.lastname: Schäfer
authors.forename: Claudia
authors.initials: C
authors.lastname: Beimfohr
authors.forename: Peter
authors.initials: P
authors.lastname: Kämpfer
mesh_terms: D000192:Actinomycetales / Q000145:classification* / Q000302:isolation & purification*; D000391:Air Microbiology; D000393:Air Pollutants / Q000032:analysis / Q000145:classification; D015374:Biosensing Techniques; D015169:Colony Count, Microbial / Q000295:instrumentation / Q000379:methods; D017404:In Situ Hybridization, Fluorescence / Q000295:instrumentation / Q000379:methods*; D008848:Micromonosporaceae / Q000145:classification / Q000302:isolation & purification; D015347:RNA Probes; D012336:RNA, Ribosomal, 16S; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D000393:Air Pollutants; D015347:RNA Probes; D012336:RNA, Ribosomal, 16S