PURPOSE: To evaluate the polymerase chain reaction (PCR) and sequencing-based ribosomal deoxyribonucleic acid (rDNA) typing technique for rapid and sensitive diagnosis of fungal keratitis. DESIGN: Laboratory investigation. METHODS: The present study included culture-based methods along with PCR and ITS2/5.8S rDNA sequencing to evaluate fungal infections in 32 corneal scrapings from patients with suspected mycotic keratitis. RESULTS: Good correlation was found between the culture and PCR rDNA sequencing-based methods. We also identified fungal strains that were noncultivable in routine culture medium and fungal strains, which have been reported scarcely in previous reports from eastern Indian studies. CONCLUSIONS: The PCR and rDNA sequencing-based approaches may be useful alternatives to culture-based techniques for the rapid and sensitive diagnosis of fungal keratitis. We found that clinically significant pathogenic fungal strains can be detected and differentiated by this method in a short period. This method therefore is useful for the early treatment of fungal infection.
PURPOSE: To evaluate the polymerase chain reaction (PCR) and sequencing-based ribosomal deoxyribonucleic acid (rDNA) typing technique for rapid and sensitive diagnosis of fungal keratitis. DESIGN: Laboratory investigation. METHODS: The present study included culture-based methods along with PCR and ITS2/5.8S rDNA sequencing to evaluate fungal infections in 32 corneal scrapings from patients with suspected mycotic keratitis. RESULTS: Good correlation was found between the culture and PCR rDNA sequencing-based methods. We also identified fungal strains that were noncultivable in routine culture medium and fungal strains, which have been reported scarcely in previous reports from eastern Indian studies. CONCLUSIONS: The PCR and rDNA sequencing-based approaches may be useful alternatives to culture-based techniques for the rapid and sensitive diagnosis of fungal keratitis. We found that clinically significant pathogenic fungal strains can be detected and differentiated by this method in a short period. This method therefore is useful for the early treatment of fungal infection.