Evaluation of polymerase chain reaction-based ribosomal DNA sequencing technique for the diagnosis of mycotic keratitis.
OBJECTIVE
To evaluate the polymerase chain reaction (PCR) and sequencing-based ribosomal deoxyribonucleic acid (rDNA) typing technique for rapid and sensitive diagnosis of fungal keratitis.


METHODS
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.
Ghosh
Abhrajyoti
A
Dr. B C Guha Centre for Genetic Engineering and Biotechnology, University of Calcutta, Kolkata, India.
Basu
Sayan
S
Datta
Himadri
H
Chattopadhyay
Dhrubajyoti
D
D000293:Adolescent; D000328:Adult; D000368:Aged; D003315:Cornea / Q000382:microbiology; D003320:Corneal Ulcer / Q000175:diagnosis* / Q000382:microbiology; D004271:DNA, Fungal / Q000032:analysis; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D015821:Eye Infections, Fungal / Q000175:diagnosis* / Q000382:microbiology; D005260:Female; D005658:Fungi / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D008875:Middle Aged; D016533:Mycological Typing Techniques; D009181:Mycoses / Q000175:diagnosis* / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012340:RNA, Ribosomal, 5.8S / Q000235:genetics*; D017422:Sequence Analysis, DNA
D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer; D012340:RNA, Ribosomal, 5.8S