2003 Fiscal Year Final Research Report Summary
Strategy for the treatment of herpetic keratitis according to the quantitative PCR method
| Project/Area Number |
14370565
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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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 | Kinki University |
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Principal Investigator |
SHIMOMURA Yoshikazu Kinki University School of Medicine, Department of Ophthalmology, Professor and Chairman, 医学部, 教授 (20162737)
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| Co-Investigator(Kenkyū-buntansha) |
HIBINO Tsuyoshi Kinki University School of Medicine, Department of Ophthalmology, Assistant professor, 医学部, 講師 (60288917)
ABE Kosuke Kinki University School of Medicine, Department of Ophthalmology, Assistant professor, 医学部, 講師 (00222646)
FUKUDA Masahiko Kinki University School of Medicine, Department of Ophthalmology, Assistant professor, 医学部, 講師 (40218938)
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| Project Period (FY) |
2002 – 2003
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| Keywords | Heipetic Keratitis / real-time PCR / HSV genome / tear sample |
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| Research Abstract |
Purpose : To understand the pathogenesis of herpetic eye diseases, we quantitated herpes simplex virus(HSV) genomes in clinical samples(tear film, aqueous humor and conical tissue) obtained from various herpetic eye diseases.
Methods : We sampled tear film from various herpetic keratitis ; epithelial keratitis(HEK)(38eyes/38patients), active stromal keratitis(HSK)(22/22), silent HSK(8/8), endotheliitis(3/3), persistent epithelial defect(PED)(9/9), aqueous humor from uveitis(6/6) and host corneas obtained from the patients who had past history of herpetic keratitis(6/6). ABI PRISM^<TM>7700 Sequence Detector(PE Applied Biosystems) was used for a real-time PCR assay.
Results : This assay had a wide linear range from 1 to 10^7 copies. The values of coefficient of variation intra-and inter-assay were 3.0% and 10.1%, respectively. HSV-DNA were detected in 81.5% of HEK patients(3.9×10^6 copies/sample in an average), 59.1% of active HSK patients(8.9×10^5), none in silent HSK and endotheliitis, 88.9% of PED patients(9.2×10^4), 16.7% of uveitis(3.82×10^5 copies/ml) and 83.3% of corneas(1.9×10^4 copies/mg). The amount of HSV-DNA in HEK patients was significantly higher than that in active HSK(P<0.05). After clinical amelioration by acyclovir, the copy numbers became below the sensitivity level of this method.
Conclusions : HSV-DNA was detected in active HSK,PED in tear samples and the host corneas having past history of HSV keratitis as well as HEK. A real-time PCR is a useful method for quantitatiation of HSV genome in herpetic eye diseases.
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