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Development of sensitive detection methods for viral shedding of adenovirus vector and replication-competent adenovirus

Research Project

Project/Area Number 17590141
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Medical pharmacy
Research InstitutionNational Institute of Health Sciences

Principal Investigator

UCHIDA Eriko  National Institute of Health Sciences, Division of cellular and gene therapy products, Section Chief (80176685)

Co-Investigator(Kenkyū-buntansha) YAMAGUCHI Teruhide  National Institute of Health sciences, Division of Biological Chemistry and Biological, Head (50111117)
NAGATA Ryuji  National Institute of Health sciences, Division of cellular and gene therapy products, Senior Researcher (20370942)
Project Period (FY) 2005 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2006: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2005: ¥2,200,000 (Direct Cost: ¥2,200,000)
Keywordsgene therapy / adenovirus vector / replication-competent adenovirus / infectivity PCR / polyethyleneimineviral / vector sheddine / virus detection method / magnetofection
Research Abstract

Detection of virus/vector shedding is one of the most important issues for safety of adenovirus vector products in gene therapy clinical research. In order to detect replication-competent adenovirus (RCA) and adenovirus vector (AdV) in blood samples of patients more sensitively and rapidly, we tried to establish a detection methods for AdV and RCA in human blood samples.
First, infectivity PCR was established for the detection of RCA and AdV. In this method, 293 cells were infected with RCA or AdV samples, and amplified RCA or AdV were quantified by real-time PCR. We showed that infectivity PCR was useful for the rapid and sensitive detection of RCA and AdV. However, infectivity PCR did not work well to detect RCA and AdV in blood samples. Therefore, we then established forcibly infection method using PEI-magnetic beads and magnetic plate for highly effective infection to cells. We also establish another forcibly infection method using protein G and anti-adenovirus antibody. Using PEI-beads method, infection efficiency of AdV increased more than 100 times.
We combined infectivity PCR and PEI-beads infection for detection of RCA and AdV in blood samples. However, infection or AdV and RCA was severely inhibited by blood samples. Therefore, another approach is required to detect RCA and AdV in blood samples more sensitively.

Report

(3 results)
  • 2006 Annual Research Report   Final Research Report Summary
  • 2005 Annual Research Report
  • Research Products

    (12 results)

All 2007 Other

All Journal Article (12 results)

  • [Journal Article] 遺伝子治療薬及び細胞治療薬のウイルス安全性確保2007

    • Author(s)
      内田 恵理子, 石井(渡部) 明子, 山口 照英
    • Journal Title

      臨床とウイルス 35

      Pages: 278-290

    • NAID

      10019820271

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Regulatory Aspects of Oncolytic Virus Products2007

    • Author(s)
      Teruhide Yamaguchi, Eriko Uchida
    • Journal Title

      Current Cancer Drvg Targets 7

      Pages: 203-208

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] 日米EU医薬品規制調和国際会議遺伝子治療専門家会議の活動と遺伝子治療薬の規制における国際動向2007

    • Author(s)
      山口 照英, 内田 恵理子
    • Journal Title

      Drug Delivery System 22-6

      Pages: 651-659

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Optimization of the virus concentration method using polyethyleneimine-conjugated magnetic beads and its application to the detection of human hepatitis A,B and C viruses2007

    • Author(s)
      Eriko Uchida, Mieko Kogi, Tadashi Oshizawa, Koei Satoh, Akiko Iwata, Mitsuhiro Murata, Mikio Hikata, Teruhide Yamaguchi
    • Journal Title

      Journal of Virological Methods 143

      Pages: 95-103

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] 遺伝子治療薬開発の現状と品質・安全性確保における国際的動向2007

    • Author(s)
      内田 恵理子
    • Journal Title

      Pharmstage 7(9)

      Pages: 1-5

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Viral safety of gene therapy and cell therapy products(Japanese)2007

    • Author(s)
      Uchida, E., et. al.
    • Journal Title

      Clinical Virology 35(4)

      Pages: 278-290

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Regulatory Aspects of Oncolytic Virus Products2007

    • Author(s)
      Yamaguchi, T., Uchida, E.
    • Journal Title

      Current Cancer Drug Targets 7

      Pages: 203-208

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Quality control and safety of human gene therapy products(Japanese)2007

    • Author(s)
      Tamaguchi, T., Uchida, E.
    • Journal Title

      Drug Delivery System 22-6

      Pages: 651-659

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Optimization of the virus concentration method using polyethyleneimine-conjugated magnetic beads and its application to the detection of human hepatitis A, B and C viruses2007

    • Author(s)
      Eriko Uchida, et. al.
    • Journal Title

      Journal of Virological Methods 143

      Pages: 95-103

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] International trends for development, quality control and safety of gene therapy products(Japanese)2007

    • Author(s)
      Uchida, E.
    • Journal Title

      Pharmstage 7(9)

      Pages: 1-5

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Regulatory Aspects of Oncolytic Virus Products2007

    • Author(s)
      Teruhide Yamaguchi, Eriko Uchida
    • Journal Title

      Current Cancer Drug Targets 7

      Pages: 203-208

    • Related Report
      2006 Annual Research Report
  • [Journal Article] 遺伝子治療薬及び細胞治療薬のウイルス安全性確保

    • Author(s)
      内田恵理子 他2名
    • Journal Title

      臨床とウイルス (In press)

    • NAID

      10019820271

    • Related Report
      2006 Annual Research Report

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Published: 2005-04-01   Modified: 2016-04-21  

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