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

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.

journal_name

Int J Food Microbiol

journal_title

International journal of food microbiology

authors

Wrent P,Rivas EM,Gil de Prado E,Peinado JM,de Silóniz MI

doi

10.1016/j.ijfoodmicro.2014.10.011

subject

Has Abstract

pub_date

2015-01-16 00:00:00

pages

109-13

eissn

0168-1605

issn

1879-3460

pii

S0168-1605(14)00515-7

journal_volume

193

pub_type

杂志文章

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