title: Identification by Molecular Methods and Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry and Antifungal Susceptibility Profiles of Clinically Significant Rare Aspergillus Species in a Referral Chest Hospital in Delhi, India.
abstract: Aspergillus species cause a wide spectrum of clinical infections. Although Aspergillus fumigatus and Aspergillus flavus remain the most commonly isolated species in aspergillosis, in the last decade, rare and cryptic Aspergillus species have emerged in diverse clinical settings. The present study analyzed the distribution and in vitro antifungal susceptibility profiles of rare Aspergillus species in clinical samples from patients with suspected aspergillosis in 8 medical centers in India. Further, a matrix-assisted laser desorption ionization-time of flight mass spectrometry in-house database was developed to identify these clinically relevant Aspergillus species. β-Tubulin and calmodulin gene sequencing identified 45 rare Aspergillus isolates to the species level, except for a solitary isolate. They included 23 less common Aspergillus species belonging to 12 sections, mainly in Circumdati, Nidulantes, Flavi, Terrei, Versicolores, Aspergillus, and Nigri Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) identified only 8 (38%) of the 23 rare Aspergillus isolates to the species level. Following the creation of an in-house database with the remaining 14 species not available in the Bruker database, the MALDI-TOF MS identification rate increased to 95%. Overall, high MICs of ≥2 μg/ml were noted for amphotericin B in 29% of the rare Aspergillus species, followed by voriconazole in 20% and isavuconazole in 7%, whereas MICs of >0.5 μg/ml for posaconazole were observed in 15% of the isolates. Regarding the clinical diagnoses in 45 patients with positive rare Aspergillus species cultures, 19 (42%) were regarded to represent colonization. In the remaining 26 patients, rare Aspergillus species were the etiologic agent of invasive, chronic, and allergic bronchopulmonary aspergillosis, allergic fungal rhinosinusitis, keratitis, and mycetoma.
authors.affiliation: Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India.
authors.forename: Aradhana
authors.initials: A
authors.lastname: Masih
authors.affiliation: Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India.
authors.forename: Pradeep K
authors.initials: PK
authors.lastname: Singh
authors.affiliation: Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India.
authors.forename: Shallu
authors.initials: S
authors.lastname: Kathuria
authors.affiliation: Department of Pulmonary Medicine, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India.
authors.forename: Kshitij
authors.initials: K
authors.lastname: Agarwal
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: 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: D000935:Antifungal Agents / Q000494:pharmacology; D001230:Aspergillus / Q000145:classification* / Q000187:drug effects / Q000302:isolation & purification*; D002147:Calmodulin / Q000235:genetics; D006761:Hospitals; D006801:Humans; D007194:India; D008826:Microbial Sensitivity Tests; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D055732:Pulmonary Aspergillosis / Q000175:diagnosis* / Q000382:microbiology; D017422:Sequence Analysis, DNA; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014404:Tubulin / Q000235:genetics
chemical_list: D000935:Antifungal Agents; D002147:Calmodulin; D014404:Tubulin