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Diagnosis of Fusarium keratitis in an animal model using the polymerase chain reaction
G Alexandrakis1, S Jalali, P Gloor
1Department of Ophthalmology and Visual Science, Yale University School of Medicine, New Haven, CT 06520-8061, USA.
Aims/Background:
The purpose of this study was apply the polymerase chain reaction (PCR) to develop a sensitive, specific, and rapid test to diagnose Fusarium keratitis. Fusarium is the most common cause of fungal corneal infection in some parts of the world. It is often difficult to establish that a keratitis is due to fungal infection.
Methods:
Fusarium solani keratitis was induced in three eyes of three rabbits by injection of a suspension of the fungus into the anterior corneal stroma. In one rabbit the contralateral eye served as a control. From four to 28 days after inoculation, the corneas were scraped for culture, then scraped and swabbed for PCR analysis. The PCR was performed with primers directed against a portion of the Fusarium cutinase gene, and the presence or absence of this amplified target sequence was determined by agarose gel.
Results:
The amplified DNA sequence was detected in 25 of 28 samples from the corneas infected with Fusarium, for a sensitivity of 89%. Only three of the 14 samples from these eyes with Fusarium keratitis were positive by culture, for a sensitivity of 21%. Seven of eight control samples were negative by the PCR based test, for a specificity of 88%.
Conclusion:
This PCR based test holds promise of being an effective method of diagnosing Fusarium keratitis as well as Fusarium infections at other sites.
Insights
This study developed a polymerase chain reaction (PCR) test for diagnosing Fusarium keratitis. The PCR test demonstrated high sensitivity and specificity for detecting fungal infections in corneal samples.
Area of Science:
- Ophthalmology
- Mycology
- Molecular Biology
Background:
- Fusarium keratitis is a challenging fungal corneal infection.
- Accurate diagnosis of fungal keratitis can be difficult.
Purpose of the Study:
- To develop a sensitive, specific, and rapid polymerase chain reaction (PCR) test for diagnosing Fusarium keratitis.
- To evaluate the efficacy of PCR in detecting Fusarium DNA in corneal samples.
Main Methods:
- Fusarium solani keratitis was induced in rabbit corneas.
- Corneal samples were analyzed using both culture and PCR.
- PCR utilized primers targeting the Fusarium cutinase gene.
Main Results:
- The PCR test detected Fusarium DNA in 89% of infected corneal samples.
- Culture methods identified Fusarium in only 21% of infected samples.
- The PCR test showed 88% specificity in distinguishing Fusarium infections.
Conclusions:
- The developed PCR-based test is a promising tool for diagnosing Fusarium keratitis.
- This method could also be applied to diagnose Fusarium infections at other body sites.
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