Development a hydrolysis probe-based quantitative PCR assay for the specific detection and quantification of .

Background And Purpose: 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 than mycological methods. Regarding this, the present study aimed to develop a hydrolysis probe-based qPCR assay for the rapid, accurate identification of .

Materials And Methods: The internal transcribed spacer 2 regions in the nuclear ribosomal DNA of and other related yeasts were assayed to find a specific PCR target for . 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 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 with a detection limit of 1 CFU per reaction. Specificity was confirmed by the non-amplification of the sequences belonging to other 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=0.99; slope=-3.42).

Conclusion: The applied qPCR assay facilitated the rapid and accurate identification and quantification of emerging opportunistic . 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 .