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T. Suzuki, T. Uno, S. Aoki, Y. Kawamura, T. Ezaki, Y. Ohashi; Application of Multiplex–PCR and DNA–microarray techniques for identifying conjunctivitis–causing pathogens . Invest. Ophthalmol. Vis. Sci. 2004;45(13):1656.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: It is difficult to identify a specific pathogen from a variety of conjunctivitis–causing agents (including viruses / bacteria / fungi) at one time, because the mode of the method of detecting each pathogen is different one from another. Therefore, we designed a new diagnostic system consisting with Multiplex–PCR and DNA–microarray techniques for the purpose of screening most of the conjunctivitis–causing pathogens in a single procedure. Methods: We amplified the selective genes of representative pathogens causing conjunctivitis and a few drug–resistance genes using specific primers. For adenoviruses, we prepared a specific DNA amplicon corresponding to each serotype. We fixed desired PCR amplicon on a slide glass to make up a DNA microarray for conjunctivitis. We amplified the selective genes of the type strain of each causative agent by Multiplex–PCR, labeling with Cy3–dye. The labeled PCR amplicon was applied on a DNA microarray to confirm its specificity. In another study, we attempted to detect the causative agent from the specimens of the patient with conjunctivitis. Results: The specificity of the reaction with our new diagnostic system was confirmed for all type strains examined. We also succeeded in distinguishing the serotype of adenoviruses. In addition, against several clinical specimens of adenoviral, herpetic, chlamydial, or bacterial conjunctivitis, this diagnostic system provided the results consistent with the diagnosis as was achieved by clinical impression or confirmed by laboratory test. Conclusions: With its specificity and rapidity, combination of Multiplex–PCR and DNA–microarray techniques can be a useful diagnostic tool for conjunctivitis patients.
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