development a hydrolysis probe-based quantitative pcr assay for the specific detection and quantification of candida auris

Background and purpose: candida auris is an emerging multidrug-resistant pathogen. the identification of this species with the conventional phenotypic or biochemical mycological methods may lead to misidentification. molecular-based species-specific identification methods such as quantitative real-time polymerase chain reaction (qpcr) facilitate a more reliable identification of c. auris than mycological methods. regarding this, the present study aimed to develop a hydrolysis probe-based qpcr assay for the rapid, accurate identification of c. auris. materials and methods: the internal transcribed spacer 2 regions in the nuclear ribosomal dna of c. auris and other related yeasts were assayed to find a specific pcr target for c. auris. a 123-base-pair target was selected, and primers and a probe were designed for hydrolysis probe-based real-time pcr with taqman chemistry. ten-fold serial dilutions of c. auris ranging from 106 to 100 cfu/ml were prepared to establish a standard curve to quantify the yeast. results: the qpcr assay was able to identify and quantify c. auris with a detection limit of 1 c. auris cfu per reaction. specificity was confirmed by the non-amplification of the sequences belonging to other candida species, yeasts, molds, bacteria, or human dnas. the standard curve of the assay showed a highly significant linearity between threshold values and dilution rates (r^2=0.99; slope=−3.42). conclusion: the applied qpcr assay facilitated the rapid and accurate identification and quantification of emerging opportunistic c. auris. therefore, considering the promising test validation results, we succeeded to develop a rapid and accurate hydrolysis probe-based qpcr assay for the screening and identification of c. auris.