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Development of species-specific primers for rapid identification of Debaryomyces hansenii

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Wrent, Petra and Rivas, Eva María and Gil de Prado, Elena and Martínez Peinado, José and Siloniz Jiménez, María Isabel de (2015) Development of species-specific primers for rapid identification of Debaryomyces hansenii. International Journal of Food Microbiology, 193 . pp. 109-113. ISSN 0168-1605

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Official URL: http://www.sciencedirect.com/science/article/pii/S0168160514005157




Abstract

In this work, we developed a specific PCR assay for Debaryomyces hansenii strains that uses a putative homologous PAD1 region (729 bp) present in this yeast species as a target. The amplification of this sequence with the D. hansenii specific primer pair (DhPADF/DhPADR) was found to be a rapid, specific and an affordable method enabling identification of D. hansenii from other yeast strains. Primers were tested in almost 100 strains, 49 strains from Type Culture Collection belonging to the genus Debaryomyces and to other yeast species commonly found in foods or related genera. These primers were able to discriminate between closely related species of Debaryomyces, such as Debaryomyces fabryi and Debaryomyces subglobosus, with a 100% detection rate for D. hansenii. Also, the method was tested in 45 strains from different foods. Results confirmed the specificity of the PCR method and detected two earlier misidentifications of D. hansenii strains obtained by RFLP analysis of the 5.8S ITS rDNA region. Subsequently we confirmed by sequencing the D1/D2 domain of 26S rDNA that these strains belonged to D. fabryi. We call attention in this work to the fact that the RFLPs of the 5.8S ITS rDNA profiles of D. hansenii, D. fabryi and D. subglobosus are the same and this technique will thus lead to incorrect identifications.


Item Type:Article
Uncontrolled Keywords:Debaryomyces hansenii, Yeasts, Species-specific primers, PCR, Rapid-identification, Foods
Subjects:Medical sciences > Biology > Biotechnology
Medical sciences > Biology > Microbiology
ID Code:32727
Deposited On:30 Jul 2015 11:34
Last Modified:10 Dec 2018 15:30

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