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Molecular characterization of drug-resistant and drug-sensitive Aspergillus isolates causing infectious keratitis
- Source :
- Indian Journal of Ophthalmology, Indian Journal of Ophthalmology, Vol 59, Iss 5, Pp 373-377 (2011)
- Publication Year :
- 2011
-
Abstract
- Purpose: To study the susceptibilities of Aspergillus species against amphotericin B in infectious keratitis and to find out if drug resistance had any association with the molecular characteristics of the fungi. Materials and Methods: One hundred and sixty Aspergillus isolates from the corneal scrapings of patients with keratitis were tested for susceptibilities to amphotericin B by broth microdilution method. These included Aspergillus flavus (64 isolates), A. fumigatus (43) and A. niger (53). Fungal DNA was extracted by glass bead vertexing technique. Polymerase chain reaction (PCR) assay was standardized and used to amplify the 28S rRNA gene. Single-stranded conformational polymorphism (SSCP) of the PCR product was performed by the standard protocol. Results: Of the 160 isolates, 84 (52.5%) showed low minimum inhibitory concentration (MIC) values (≤ 1.56 μg/ml) and were designated as amphotercin B-sensitive. Similarly, 76 (47.5%) had high MICs (≥ 3.12 μg/ml) and were categorized as amphotericin B-resistant. MIC 50 and MIC 90 values ranged between 3.12-6.25 μg/ml and 3.12-12.5 μg/ml respectively. A. flavus and A. niger showed higher MIC 50 and MIC 90 values than A. fumigatus . The SSCP pattern exhibited three extra bands (150 bp, 200 bp and 250 bp each) in addition to the 260 bp amplicon. Strains (lanes 1 and 7) lacking the 150 bp band showed low MIC values (≤ 1.56 μg/ml). Conclusion: A. niger and A. flavus isolates had higher MICs compared to A. fumigatus , suggesting a high index of suspicion for amphotericin B resistance. PCR-SSCP was a good molecular tool to characterize Aspergillus phenotypes in fungal keratitis.
- Subjects :
- Antifungal Agents
analysis
polymerase chain reaction--single-stranded conformational polymorphism
Aspergillus flavus
Drug resistance
Behcet′s disease
Polymerase Chain Reaction
Graves′ ophthalmopathy
law.invention
Cornea
lcsh:Ophthalmology
law
Amphotericin B
Fungal keratitis
Assam
Polymerase chain reaction
Polymorphism, Single-Stranded Conformational
biology
Broth microdilution
Antifungal sensitivity testing
amphotericin B
Pediatric traumatic cataract
corneal ulcer
Aspergillus
fungal keratitis
Original Article
Eye Infections, Fungal
square-edge intraocular lens
medicine.drug
posterior capsule opacification
HLA-B51
Microbiology
Minimum inhibitory concentration
visual axis opacification
Drug Resistance, Fungal
retinitis pigmentosa
medicine
unilateral
Aspergillosis
Humans
fusarium
Keratitis
optical coherence tomography
mycotic keratitis
RNA, Fungal
primary posterior capsulotomy
thyroid eye disease
medicine.disease
biology.organism_classification
macular hole
Ophthalmology
lcsh:RE1-994
principal components′
Multifocal electroretinogram
Subjects
Details
- ISSN :
- 19983689
- Volume :
- 59
- Issue :
- 5
- Database :
- OpenAIRE
- Journal :
- Indian journal of ophthalmology
- Accession number :
- edsair.doi.dedup.....db435156f5ed36049ea4c0d4700b4853