TY - JOUR
T1 - Identification of yoghurt-spoiling yeasts with 18S rRNA-targeted oligonucleotide probes
AU - Kosse, Dagmar
AU - Seiler, Herbert
AU - Amann, Rudolf
AU - Ludwig, Wolfgang
AU - Scherer, Siegfried
N1 - Funding Information:
Acknowledgements We thank N. VALDES-STAUBER for valuable discussions and helpful support in the laboratory. Thanks is also due to I. OSTENRIEDER for excellent laboratory cooperation. Part of this work was supported by the Forschungskreis der Ernahrungsindustrie (FEll AiF), Bonn.
PY - 1997/8
Y1 - 1997/8
N2 - 18S rRNA-targeted oligonucleotide probes were designed for rapid and reliable identification of yeasts involved in spoilage of dairy products. Candida parapsilosis, Candida glabrata, Clavispora lusitaniae, Debaryomyces hansenii, Dekkera bruxellensis, Hanseniaspora uvarum, Pichia anomala, Pichia membranaefaciens, Rhodotorula glutinis, Saccharomyces cerevisiae and Saccharomycopsis capsularis were identified by dot blot hybridization assays using species-specific, digoxygenin-labeled probes which were deduced from comparative analysis of highly variable regions of 18S rRNA. The eucarya-specific probe EUK516 gave intense signals detected by epifluorescense microscopy when hybridized in situ to all yeast species tested. Whole cell hybridization experiments revealed that the 3'-end of the target molecule is a suitable site for fluorescently labeled species-specific nucleic acid probes which detect S. cerevisiae, P. anomala, D. hansenii and D. bruxellensis in situ. Other variable regions of the 18S rRNA tested for species-specific probes apparently were not accessible to in situ hybridization. S. cerevisiae and P. anomala were detected in yoghurt using the fluorescently labeled probes Sce1711 and Pan1710.
AB - 18S rRNA-targeted oligonucleotide probes were designed for rapid and reliable identification of yeasts involved in spoilage of dairy products. Candida parapsilosis, Candida glabrata, Clavispora lusitaniae, Debaryomyces hansenii, Dekkera bruxellensis, Hanseniaspora uvarum, Pichia anomala, Pichia membranaefaciens, Rhodotorula glutinis, Saccharomyces cerevisiae and Saccharomycopsis capsularis were identified by dot blot hybridization assays using species-specific, digoxygenin-labeled probes which were deduced from comparative analysis of highly variable regions of 18S rRNA. The eucarya-specific probe EUK516 gave intense signals detected by epifluorescense microscopy when hybridized in situ to all yeast species tested. Whole cell hybridization experiments revealed that the 3'-end of the target molecule is a suitable site for fluorescently labeled species-specific nucleic acid probes which detect S. cerevisiae, P. anomala, D. hansenii and D. bruxellensis in situ. Other variable regions of the 18S rRNA tested for species-specific probes apparently were not accessible to in situ hybridization. S. cerevisiae and P. anomala were detected in yoghurt using the fluorescently labeled probes Sce1711 and Pan1710.
KW - 18S rRNA
KW - Dairy products
KW - Dot blot hybridization
KW - Identification
KW - In situ hybridization
KW - Oligonucleotide probes
KW - Yeasts
UR - http://www.scopus.com/inward/record.url?scp=0030858069&partnerID=8YFLogxK
U2 - 10.1016/S0723-2020(97)80016-1
DO - 10.1016/S0723-2020(97)80016-1
M3 - Article
AN - SCOPUS:0030858069
SN - 0723-2020
VL - 20
SP - 468
EP - 480
JO - Systematic and Applied Microbiology
JF - Systematic and Applied Microbiology
IS - 3
ER -