2004 Fiscal Year Final Research Report Summary
Molecular analysis of virus genomes/transcript in the intraocular fluid samples during viral uveitis
| Project/Area Number |
14571687
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Ophthalmology
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| Research Institution | Keio University |
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Principal Investigator |
ANDO Yasutaka Keio University, School of Medicine, Department of Ophthalmology, Assistant Professor, 医学部, 講師 (90193119)
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| Co-Investigator(Kenkyū-buntansha) |
IWASAKI Takuya Nagasaki University, Institute of Tropical Medicine, Professor, 熱帯医学研究所, 教授 (90146027)
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| Project Period (FY) |
2002 – 2004
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| Keywords | viral uveitis / cytomegalovirus retinitis / acute retinal necrosis / intraocular fluid / real-time PCR / LAMP |
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| Research Abstract |
In this study we investigated the quantitative diagnostic methods useful for rapid diagnosis and treatment of viral diseases of the eye. Especially, we focused on the cytomegalovirus (CMV) retinitis and acute retinal necrosis. Using the real time polymerase chain reaction (PCR) we determined the amount of the viral genomes of human CMV and varicella-zoster virus (VZV) in fluid samples aspirated from the eyeballs. We found the positive correlation between the copy numbers of the viral genomes and the lesion activity. These copy numbers became rapidly decreased after the antiviral treatment. In one patient with immune recovery uveitis after CMV retinitis, the copy number of human CMV genome in the vitreous fluid was less than the level of detection, but the high amount of viral antigens could he detected. As next step we further studies a more prompt, specific and simple diagnostic method using a recently developed loop-mediated isothermal amplification (LAMP). Using this LAMP method we determined the viral genomes of VZV in patients with acute retinal necrosis and found to be reliable to do this analysis. The result by this method was well correlated with that by real-time PCR, but the sensitively was less than that by real time PCR. In addition, we could apply the samples without DNA extraction for LAMP analysis except one specimen. In conclusion, LAMP method is semi-quantitative but simple and rapid. From this conclusion, this method will he valuable for the clinical application for diagnosis of viral ocular diseases.
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