Kefir is a product elaborated from the symbiotic fermentation of different microorganisms. The Kluyveromyces and Saccharomyces genera are the major representatives of the yeasts found in kefir microbiota. The only pobiotic yeast commercialized as an oral medication, is the Saccharomyces boulardii. The present work involved the microbiological quality examination of six kefir samples in the city of Santa Maria/RS, the yeasts isolation present in the samples and the identification of them by PCR (Polymerase chain reaction). Then, their probiotic potential was evaluated by in vitro technique. After that, microbiological analysis confirmed that kefir samples were suitable for consumption once the microbiological quality was established. Nineteen yeast strains were isolated from six different kefir samples; it was identified, by PCR analysis, but only three species were identified from these microorganisms in the present article: Saccharomyces cerevisiae, Hanseniospora uvarum and Kazachstania unispora. Nevertheless, by simulating the passage of isolated strains through the gastrointestinal environment, it was observed that they could not be considered probiotics. The results indicate that, in an isolated way, the yeast presents in kefir samples, in the city of Santa Maria, RS, can´t be considered probiotics according to the tests performed.