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KINETICS OF OCULAR HSV-1 -QUANTITATION OF PCR-

Research Project

Project/Area Number 07457412
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Ophthalmology
Research InstitutionOsaka University

Principal Investigator

SHIMOMURA Yoshikazu  Osaka University, Ophthalmology, Associate Professor, 医学部, 助教授 (20162737)

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)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
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)
KeywordsHSV-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
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.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (9 results)

All Other

All Publications (9 results)

  • [Publications] 下村 嘉一: "眼感染症の新しい診断法" あたらしい眼科. 14・2. 193-197 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Mori Y, et al: "Detection of HSV mRNA using revarse transcription polymerase chain reaction for diagnosis in murine herpetic keratitls model" Jpn J Ophthamol. 42・1. 8-11 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Inoue T, et al: "Corneal infiltration and CMV retinitis in apatient with AIDS" Cornea. (aceepted).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Shimomura Y: "Curent Opinions in the Kyoto Cornea Club" Kugler Publications,Netherlands, 5 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Shimomura Y: "Usefulness of polymerase chain reaction technology in identifying infections agents causing ocular diseases" Atarashii Ganka. 14-2. 193-197 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Mori Y,Inoue Y,Shimomura Y,Kase T,Tano Y: "Detection of HSV mRNA using reverse transcription-polymerase chain reaction for diagnosis in murine herpetic keratitis model" Jpn J Ophthalmol. 42-1. 8-11 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Inoue T,Hayashi K,Omoto T,Kosaki R,Higaki S,Shimomura Y: "Corneal infiltration and CMV retinitis in a patient with AIDS" Gornea. (accepted).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Shimomura Y: Herpes simplex virus latency in the cornea(Kinoshita S and Ohashi Y,ed) Current Opinions in the Kyoto Cornea Club. Kugler Publications, Netherlands, 63-67 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 下村 嘉一: "眼感染症の新しい診断法" あたらしい眼科. 14・2. 193-197 (1997)

    • Related Report
      1996 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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