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Summary 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. |
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