Random amplified ribosomal DNA restriction analysis for rapid identification of thermophilic Actinomycete-like bacteria involved in hypersensitivity pneumonitis |
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| Authors: | Harvey I Cormier Y Beaulieu C Akimov V N Mériaux A Duchaine C |
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| Affiliation: | 1. Unité de recherche, Centre de recherche, Hôpital Laval, Institut universitaire de cardiologie et de pneumologie de l''Université Laval, Ste-Foy, Québec, Canada;2. Département de biologie, Université de Sherbrooke, QC, Canada;1. Department of Gastroenterology, Second Affiliated Hospital of Qiqihar Medical University, Qiqihar, 161000, China;2. Department of General Medicine, Second Affiliated Hospital of Qiqihar Medical University, Qiqihar, 161000, China;3. Department of Respiratory, Second Affiliated Hospital of Qiqihar Medical University, Qiqihar, 161000, China;4. Department of Gastroenterology, East Hospital of Shanghai, 200000, Shanghai, China;1. Department of Biotechnology, Sector 14, Panjab University, Chandigarh 160014, India;2. National Centre for Human Genome Studies and Research, Panjab University, Chandigarh 160014, India;1. Department of Chemical Engineering, Osaka Prefecture University, 1-1 Gakuen-cho, Naka-ku, Sakai, Osaka 599-8531, Japan;2. Department of Biological Science, Graduate School of Science, Osaka Prefecture University, 1-2 Gakuen-cho, Naka-ku, Sakai, Osaka 599-8570, Japan;1. Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens, 9 Iroon Polytechniou Str., Zografou Campus, 15780, Athens, Greece;2. Environment and Quality of Life Center, School of Chemical Engineering, National Technical University of Athens, 9 Iroon Polytechniou Str., Zografou Campus, 15780, Athens, Greece |
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| Abstract: | Hypersensitivity pneumonitis (HP) is a pulmonary disease characterised by inflammation that can be caused by, amongst other substances, a subset of 4 thermophilic mycelial bacteria: Saccharopolyspora rectivirgula, Saccharomonospora viridis, Thermoactinomyces sacchari, and Thermoactinomyces vulgaris. Air sampling analyses in highly contaminated environments are often performed to evaluate exposure to these species which are difficult and fastidious to identify by conventional techniques. The aim of this study was to use amplified ribosomal DNA restriction analysis (ARDRA) to develop a method of identification for those thermophilic organisms that would be more rapid and simple. Strains of these 4 species were obtained from the American type culture collection (ATCC) and were characterized using biochemical tests and ARDRA patterns obtained on their partial-lenght amplified 16S rDNAs. To validate this approach, ARDRA with two restriction enzymes, TaqI and HhaI, was applied to 49 thermophilic actinomycete-like strains from environmental samples (sawmills). The results obtained show that combining some cultural characteristics and biochemical tests, such as xanthine or hypoxanthine decomposition, growth in the presence of NaCl, lysozyme or novobiocin, and spore resistance over 100 degrees C provide a rough identification and selection of the genera of interest. Consequently, target species could be confirmed by digestion of partial-lenght 16S rDNA with the use of Taql and HhaI restriction enzymes that gave specific restriction patterns. ARDRA analyses on the 49 environmental actinomycete-like organisms revealed the presence of 8 Saccharopolyspora rectivirgula, 2 Saccharomonospora viridis, and 15 Thermoactinomyces vulgaris strains, the other strains had restriction patterns different than those of the species of interest. Results of the present study will be applicable to other potential HP environments such as dairy barns, peat bogs and compost plants. |
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| Keywords: | ARDRA thermophilic actinomycetes hypersensitivity pneumonitis environmental samples |
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