| Project/Area Number |
07457412
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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 | Osaka University |
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Principal Investigator |
SHIMOMURA Yoshikazu Osaka University, Ophthalmology, Associate Professor, 医学部, 助教授 (20162737)
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| Co-Investigator(Kenkyū-buntansha) |
HAYASHI Kozaburou Kobe Institute of Health, Virology, Director, 所長
WATANABE Hitoshi Osaka University, Ophthalmology, Research Associate, 医学部, 助手 (60252673)
KIRITOSHI Akira Osaka University, Ophthalmology, Assistant Research, 医学部, 講師 (10225104)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥4,900,000 (Direct Cost: ¥4,900,000)
Fiscal Year 1996: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1995: ¥4,000,000 (Direct Cost: ¥4,000,000)
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| Keywords | HSV-1 / PCR / QUANTITATION / keratitis / mouse / 単純ヘルペスウイルス / 培養法 / 眼局所 / 三叉神経節 |
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| Research Abstract |
1.DNA quantitation by single PCR in mouse HSV-1 keratitis model
We determined that 0.2 ng of DNA,which was equal to 400 PFU,could be detected by single PCR.Investigation in changes of virus titlation and DNA using mouse HSV-1 keratitis model showed that virus titlations in lacrimal glands could be detected more than 10^2 PFU,but were less than those in eye balls or trigeminal ganglia. DNA could not be detected in lacrimal glands or trigeminal ganglia on day 2 postinfection (PI), but detected on day 4 PI and decreased on day 6 PI not to be detected on day 8 PI.DNA could be detected in eye balls on day 2 PI to decrease gradually. These results showed that competitive PCR should be utilized for DNA quantitation instead of single PCR.
2.DNA quantitation by competitive PCR
Numbers of HSV-1 genome copy increased proportionately between 21 and 25 cycles using only competitor, while they increased proportionately between 21 and 27 cycles using only HSV-1 genome. These results showed that DNA could be quantitate between 21 and 25 cycles even if doses of HSV-1 genomes changed.
When competitive PCR was performed 25 cycles using dilution of HSV-1 genome and competitor as a template, 780 pg of HSV-1 genome was found to be equal to 10^7 copy of competitor.
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