Development of sensitive detection methods for viral shedding of adenovirus vector and replication-competent adenovirus
| Project/Area Number |
17590141
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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 |
Medical pharmacy
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| Research Institution | National Institute of Health Sciences |
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Principal Investigator |
UCHIDA Eriko National Institute of Health Sciences, Division of cellular and gene therapy products, Section Chief (80176685)
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| 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)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| 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)
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| Keywords | gene therapy / adenovirus vector / replication-competent adenovirus / infectivity PCR / polyethyleneimineviral / vector sheddine / virus detection method / magnetofection |
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| 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.
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Report
(3 results)
Research Products
(12 results)
