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
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 ty...
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| Published in: | Clinical microbiology and infection Vol. 22; no. 3; pp. 277.e1 - 277.e9 |
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| Main Authors: | , , , , , , , , , |
| Format: | Journal Article |
| Language: | English |
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01.03.2016
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| ISSN: | 1469-0691 |
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| 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. |
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| AbstractList | 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. |
| Author | Singh, A Kumar, A Kumar Singh, P Chowdhary, A Prakash, A Govender, N P Colombo, A L Meis, J F Hagen, F Sharma, C |
| Author_xml | – sequence: 1 givenname: A surname: Prakash fullname: Prakash, A organization: Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India – sequence: 2 givenname: C surname: Sharma fullname: Sharma, C organization: Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India – sequence: 3 givenname: A surname: Singh fullname: Singh, A organization: Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India – sequence: 4 givenname: P surname: Kumar Singh fullname: Kumar Singh, P organization: Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India – sequence: 5 givenname: A surname: Kumar fullname: Kumar, A organization: Department of Microbiology, Amrita Institute of Medical Sciences, Ponekara, Kochi, India – sequence: 6 givenname: F surname: Hagen fullname: Hagen, F organization: Department of Medical Microbiology and Infectious Diseases, Canisius-Wilhelmina Hospital, Nijmegen, The Netherlands – sequence: 7 givenname: N P surname: Govender fullname: Govender, N P organization: National Institute for Communicable Diseases-Centre for Opportunistic, Tropical and Hospital Infections, Johannesburg, South Africa – sequence: 8 givenname: A L surname: Colombo fullname: Colombo, A L organization: Laboratório Especial de Micologia, Disciplina de Infectologia, Escola paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP, Brazil – sequence: 9 givenname: J F surname: Meis fullname: Meis, J F organization: Department of Medical Microbiology and Infectious Diseases, Canisius-Wilhelmina Hospital, Nijmegen, The Netherlands; Department of Medical Microbiology, Radboudumc, Nijmegen, The Netherlands – sequence: 10 givenname: A surname: Chowdhary fullname: Chowdhary, A email: dranuradha@hotmail.com organization: Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India. Electronic address: dranuradha@hotmail.com |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/26548511$$D View this record in MEDLINE/PubMed |
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| Keywords | Candida auris Amplified fragment length polymorphism D1/D2 internal transcribed spacer RPB2 RPB1 matrix-assisted laser desorption ionization time-of-flight mass spectrometry |
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| SubjectTerms | Amplified Fragment Length Polymorphism Analysis Antifungal Agents - pharmacology Candida - classification Candida - drug effects Candida - genetics Candida - isolation & purification Candidiasis - epidemiology Candidiasis - microbiology DNA, Ribosomal Spacer Genes, Fungal Genotype Humans Microbial Sensitivity Tests Multilocus Sequence Typing Phenotype Phylogeny Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization |
| 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 |
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