Identification of yeasts by RFLP analysis of the 5.8S rRNA gene and the two ribosomal internal transcribed spacers

TLDR

Yeast identification has traditionally relied on morphological, physiological, and biochemical traits, while rapid kits exist mainly for clinical use and recent molecular techniques lack a comprehensive database for many species. The study aimed to identify 132 yeast species across 25 genera using restriction patterns of the ITS1/ITS2 and 5.8S rRNA gene region. Restriction patterns were generated by digesting PCR products with endonucleases CfoI, HaeIII, and HinfI to produce species‑specific profiles. The resulting restriction patterns yielded unique profiles for each species, supporting the method as a rapid and easy routine identification tool.

Abstract

The identification and classification of yeasts have traditionally been based morphological, physiological and biochemical traits. Various kits have been developed as rapid systems for yeast identification, but mostly for clinical diagnosis. In recent years, different molecular biology techniques have been developed for yeast identification, but there is no available database to identify a large number of species. In the present study, the restriction patterns generated from the region spanning the internal transcribed spacer (ITS1 and ITS2) and the 5.8S rRNA gene were used to identify a total of 132 yeast species belonging to 25 different genera, including teleomorphic and anamorphic ascomycetous and basidiomycetous yeasts. In many cases, the size of the PCR products and the restriction patterns obtained with endonucleases CfoI, HaeIII and HinfI yielded a unique profile for each species. Accordingly, the use of this molecular approach is proposed as a new rapid and easy method of routine yeast identification.