title: Molecular and Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry-Based Characterization of Clinically Significant Melanized Fungi in India.
abstract: Melanized or black fungi are a heterogeneous group of fungi causing cutaneous to systemic diseases with high mortality. These fungi are rarely reported as agents of human infections, primarily due to difficulties in their classical identification. In this study, we examined, using molecular methods and matrix-assisted laser desorption ioniazation-time of flight mass spectrometry (MALDI-TOF MS), the diversity of melanized fungi (MF) isolated from patients in 19 medical centers in India. Overall, during a 4-year period, 718 (5.3%) clinical specimens yielded MF. Of these, 72 (10%) isolates had clinical significance and were identified primarily by sequencing the internal transcribed spacer and large subunit (LSU) regions. MF represented 21 genera comprising 29 species, the majority of them belonging to the orders Pleosporales (50%) and Chaetothyriales (22%). Among the 29 fungal species identified in this study, only 6 (20%) species were identified by the MALDI-TOF MS due to the limited commercial database of Bruker Daltonics for MF. However, a 100% identification rate of 20 additional species identified in this study was obtained by constructing an in-house database using 24- to 96-h-old liquid cultures. Further, the CLSI broth microdilution method revealed low MICs for posaconazole (≤1 μg/ml) and voriconazole (≤2 μg/ml) in 96% and 95% of isolates, respectively. Skin/subcutaneous and sino-nasal and pulmonary phaeohyphomycosis due to MF were diagnosed in 21.4% (n= 15) and 28.5% (n= 20) of cases. Also, 10% of patients had central nervous system involvement (n= 7), and 3 cases of fungal osteomyelitis due toCladophialophora bantianaandCorynesporaspp. were observed.
authors.affiliation: Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India.
authors.forename: Ashutosh
authors.initials: A
authors.lastname: Singh
authors.affiliation: Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India.
authors.forename: Pradeep Kumar
authors.initials: PK
authors.lastname: Singh
authors.affiliation: Department of Microbiology, Amrita Institute of Medical Sciences, Kochi, India.
authors.forename: Anil
authors.initials: A
authors.lastname: Kumar
authors.affiliation: Department of Microbiology, Government Medical College Hospital, Chandigarh, India.
authors.forename: Jagdish
authors.initials: J
authors.lastname: Chander
authors.affiliation: Department of Pathology, Vardhman Mahavir Medical College and Safdarjung Hospital, Guru Gobind Singh Indraprastha University, New Delhi, India.
authors.forename: Geetika
authors.initials: G
authors.lastname: Khanna
authors.affiliation: Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India.
authors.forename: Pradip
authors.initials: P
authors.lastname: Roy
authors.affiliation: Department of Medical Microbiology and Infectious Diseases, Canisius-Wilhelmina Hospital, Nijmegen, The Netherlands.|Centre of Expertise in Mycology Radboudumc/CWZ, Nijmegen, The Netherlands.
authors.forename: Jacques F
authors.identifier: 0000-0003-3253-6080
authors.initials: JF
authors.lastname: Meis
authors.affiliation: Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India dranuradha@hotmail.com.
authors.forename: Anuradha
authors.identifier: 0000-0002-2028-7462
authors.initials: A
authors.lastname: Chowdhary
mesh_terms: D000368:Aged; D000935:Antifungal Agents / Q000494:pharmacology; D044822:Biodiversity; D002648:Child; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics; D005658:Fungi / Q000737:chemistry / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007194:India; D008297:Male; D008826:Microbial Sensitivity Tests; D008828:Microbiological Techniques / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009181:Mycoses / Q000175:diagnosis*; D017422:Sequence Analysis, DNA; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D000935:Antifungal Agents; D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer
keywords: India; MALDI-TOF MS; antifungal resistance; melanized fungi; phaeohyphomycosis