Identification of Candida auris by PCR and assessment of biofilm formation by crystal violet assay
Parul Singh, Smriti Srivastava, Rajesh Malhotra, Purva Mathur
Abstract
Introduction Candida auris is a notorious pathogen capable of forming biofilms on devices as well as host tissues, often culminating in infections. We evaluated characteristics of infections and the methods to diagnose C. auris over a period of three years in a tertiary care hospital. Methods Patients admitted between 2018 and 2020, who had candidemia due to C. auris were included in the study. Identification was performed using HiCrome™ Candida Differential Agar, Vitek 2 (BioMérieux, Inc., Marcy-l’Etoile, France) and MALDI-TOF, Vitek-MS. Identification was confirmed by detection of rDNA region covering part of 5.8S, entire of ITS2, and part of 28S by polymerase chain reaction (PCR). Biofilm formation was assessed by crystal violet staining. Results Presence of central line and broad spectrum antimicrobials were noted in all patients whereas total parenteral nutrition was given in 82.1% of these patients. Identification by Vitek2 v8.1 correlated with MALDI-TOF MS. PCR products of length 163 bp were obtained in all isolates as visualized by agarose gel electrophoresis . The biofilm quantity measured as A 560 of the twenty-eight C. auris isolates ranged from 0.16 to 0.80 compared to C. albicans . Conclusions C. auris can be identified by PCR targeting specific rDNA region. Biofilm formation and quantification can be achieved by growing C. auris isolates in Mueller-Hinton broth over a duration of 48 h.