TY - JOUR ID - 119575 TI - Development a hydrolysis probe-based quantitative PCR assay for the specific detection and quantification of Candida auris JO - Current Medical Mycology JA - CMM LA - en SN - 2423-3439 AU - Jafarian, Hadis AU - Khodadadi, Hossein AU - Badiee, Parisa AD - Department of Medical Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran AD - Prof. Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran Y1 - 2020 PY - 2020 VL - 6 IS - 3 SP - 50 EP - 56 KW - Candida auris KW - Quantification KW - Real-Time PCR DO - 10.18502/cmm.6.3.4665 N2 - 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 (R2=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.   UR - https://cmm.mazums.ac.ir/article_119575.html L1 - https://cmm.mazums.ac.ir/article_119575_a19b4642f6765cfb2292e723b76b6818.pdf ER -