Fusarium Keratitis From a Comprehensive Eye Health Care Facility in South India: Molecular Characterization by MALDI-TOF Versus Polymerase Chain Reaction Sequencing, Species Complex Distribution, and Clinical Correlation

Author:

Bagga Bhupesh1,Das Sourav2,Tawde Yamini2,Singh Shreya3,Shaw Tushar4,Sharma Savitri5,Ghosh Anup2ORCID

Affiliation:

1. Department of Cornea and Anterior Segment, L. V. Prasad Eye Institute, Hyderabad, Telangana, India;

2. Department of Medical Microbiology (Mycology Division), Post Graduate Institute of Medical Education and Research, Chandigarh, India;

3. Department of Microbiology, Dr B. R. Ambedkar Institute of Medical Sciences (AIMS Mohali), Chandigarh, India;

4. Department of Life and Allied Health Sciences, Ramaiah University of Applied Sciences, Bangalore, Karnataka, India; and

5. Jhaveri Microbiology Centre, L V Prasad Eye Institute, Hyderabad, Telangana, India.

Abstract

Purpose: Fusarium keratitis possesses significant diagnostic and therapeutic challenges. Medically relevant Fusaria belong to various species complexes and show prominent differences in their antifungal susceptibility profile which may influence the clinical outcome. Rapid diagnostic methods are warranted for precise identification of species complexes for prompt initiation of correct antifungals. The aim of the study was to compare between matrix-assisted laser desorption/ionization–time of flight (MALDI-TOF) and polymerase chain reaction sequencing for correct species-level identification and to analyze the clinical outcome among different Fusarium species complexes. Methods: Twenty-nine culture-proven Fusarium keratitis cases were included in this study. A phylogenetic tree was constructed after TEF1α gene sequencing and isolates were subjected to MALDI-TOF MS, followed by database expansion and identification. Clinical outcome and risk association among species complexes were analyzed retrospectively. Results: Maximum likelihood phylogeny categorized 68.9% isolates as Fusarium solani species complex (FSSC), 17.2% as Fusarium dimerum species complex (FDSC), followed by 13.7% as Fusarium fujikuroi species complex (FFSC). With extended database, MALDI-TOF MS could correctly speciate 96.5% (28/29) isolates. Previous antibiotic usage (P = 0.034) and preoperative antifungal treatment with natamycin, voriconazole, or ketoconazole (P = 0.025) were significantly higher in the FSSC group. The patients in the FFSC group had a significantly longer duration of symptoms at the time of clinical presentation to the clinic (15 days vs. 5 days, P = 0.030). Among 11 patients with a clinically poor outcome, 9 (31%) had FSSC infection. Conclusions: Patients infected with the FSSC had more aggressive infection with poor prognosis. MALDI-TOF MS can serve as the best alternative method to conventional molecular identification with reduced turnaround time, which may help the ophthalmologists to consider the appropriate antifungals or early surgical intervention for improved outcome.

Publisher

Ovid Technologies (Wolters Kluwer Health)

Subject

Ophthalmology

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