G Alexandrakis1, S Jalali, P Gloor. 1. Department of Ophthalmology and Visual Science, Yale University School of Medicine, New Haven, CT 06520-8061, USA.
Abstract
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.
AIMS/ BACKGROUND: The purpose of this study was apply the polymerase chain reaction (PCR) to develop a sensitive, specific, and rapid test to diagnose Fusariumkeratitis. 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 solanikeratitis 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 Fusariumkeratitis 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 Fusariumkeratitis as well as Fusarium infections at other sites.
Authors: Mohamed Abou Shousha; Andrea Rachelle C Santos; Rafael A Oechsler; Alfonso Iovieno; Jorge Maestre-Mesa; Marco Ruggeri; Jose J Echegaray; Sander R Dubovy; Victor L Perez; Darlene Miller; Eduardo C Alfonso; M Livia Bajenaru Journal: Mol Vis Date: 2013-12-27 Impact factor: 2.367