title: Evidence of genotypic diversity among Candida auris isolates by multilocus sequence typing, matrix-assisted laser desorption ionization time-of-flight mass spectrometry and amplified fragment length polymorphism.
abstract: Candida auris is a multidrug-resistant nosocomial bloodstream pathogen that has been reported from Asian countries and South Africa. Herein, we studied the population structure and genetic relatedness among 104 global C. auris isolates from India, South Africa and Brazil using multilocus sequence typing (MLST), amplified fragment length polymorphism (AFLP) fingerprinting and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). RPB1, RPB2 and internal transcribed spacer (ITS) and D1/D2 regions of the ribosomal DNA were sequenced for MLST. Further, genetic variation and proteomic assessment was carried out using AFLP and MALDI-TOF MS, respectively. Both MLST and AFLP typing clearly demarcated two major clusters comprising Indian and Brazilian isolates. However, the South African isolates were randomly distributed, suggesting different genotypes. MALDI-TOF MS spectral profiling also revealed evidence of geographical clustering but did not correlate fully with the genotyping methods. Notably, overall the population structure of C. auris showed evidence of geographical clustering by all the three techniques analysed. Antifungal susceptibility testing by the CLSI microbroth dilution method revealed that fluconazole had limited activity against 87% of isolates (MIC90, 64 mg/L). Also, MIC90 of AMB was 4 mg/L. Candida auris is emerging as an important yeast pathogen globally and requires reproducible laboratory methods for identification and typing. Evaluation of MALDI-TOF MS as a typing method for this yeast is warranted.
authors.affiliation: Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India.
authors.forename: A
authors.initials: A
authors.lastname: Prakash
authors.affiliation: Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India.
authors.forename: C
authors.initials: C
authors.lastname: Sharma
authors.affiliation: Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India.
authors.forename: A
authors.initials: A
authors.lastname: Singh
authors.affiliation: Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India.
authors.forename: P
authors.initials: P
authors.lastname: Kumar Singh
authors.affiliation: Department of Microbiology, Amrita Institute of Medical Sciences, Ponekara, Kochi, India.
authors.forename: A
authors.initials: A
authors.lastname: Kumar
authors.affiliation: Department of Medical Microbiology and Infectious Diseases, Canisius-Wilhelmina Hospital, Nijmegen, The Netherlands.
authors.forename: F
authors.initials: F
authors.lastname: Hagen
authors.affiliation: National Institute for Communicable Diseases-Centre for Opportunistic, Tropical and Hospital Infections, Johannesburg, South Africa.
authors.forename: N P
authors.initials: NP
authors.lastname: Govender
authors.affiliation: Laboratório Especial de Micologia, Disciplina de Infectologia, Escola paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP, Brazil.
authors.forename: A L
authors.initials: AL
authors.lastname: Colombo
authors.affiliation: Department of Medical Microbiology and Infectious Diseases, Canisius-Wilhelmina Hospital, Nijmegen, The Netherlands; Department of Medical Microbiology, Radboudumc, Nijmegen, The Netherlands.
authors.forename: J F
authors.initials: JF
authors.lastname: Meis
authors.affiliation: Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India. Electronic address: dranuradha@hotmail.com.
authors.forename: A
authors.initials: A
authors.lastname: Chowdhary
mesh_terms: D054458:Amplified Fragment Length Polymorphism Analysis; D000935:Antifungal Agents / Q000494:pharmacology; D002175:Candida / Q000145:classification* / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D002177:Candidiasis / Q000453:epidemiology / Q000382:microbiology*; D021903:DNA, Ribosomal Spacer; D005800:Genes, Fungal; D005838:Genotype; D006801:Humans; D008826:Microbial Sensitivity Tests; D058885:Multilocus Sequence Typing; D010641:Phenotype; D010802:Phylogeny; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000935:Antifungal Agents; D021903:DNA, Ribosomal Spacer
keywords: Amplified fragment length polymorphism; Candida auris; D1/D2; RPB1; RPB2; internal transcribed spacer; matrix-assisted laser desorption ionization time-of-flight mass spectrometry