1991 Fiscal Year Final Research Report Summary
Detection of hepatitis C virus RNA by sensitive PCR method
| Project/Area Number |
02807072
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Gastroenterology
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| Research Institution | Chiba University |
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Principal Investigator |
YOKOSUKA Osamu Chiba University, assistant, 医学部, 助手 (90182691)
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| Co-Investigator(Kenkyū-buntansha) |
IMAZEKI Fumio Chiba University, assistant, 医学部, 助手 (40223325)
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| Project Period (FY) |
1990 – 1991
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| Keywords | Polymerase chain reaction / Hepatitis C virus / HCV RNA / Synthetic DNA / Primer / Acute hepatitis / Chronic hepatitis / Interferon |
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| Research Abstract |
Non-A, non-B hepatitis virus is a major etiologic agent of chronic liver disease in Japan. Recently, M. Houghton et al. mleculary cloned the hepatitis C virus(HCV)nucleic acid and reported its genomic sequence. An assay system for detection of anti-HCV antibody was also established. However, detection method for HCV antigen or HCV nucleic acid have not been established. The reason might probably be because of small amount of the virus in spechmns. Here we tried to establish a sensitive method for detection of HCV RNA by applying polymerase chain reaction(PCR)which is a powerful method to detect small amount of nucleic acid. To detect the HCV RNA, we extracted the nucleic acid from patients samples(blood and liver tissue), then reversetranscribed the nucleic acid to cDNA, then amplified with PCR method using synthetic oligonucleotide primers. The primes used were synthesized based on the HCV sequence and flaviviral sequences. With this RT-PCR rmthod we detected the HCV RNA in the sera from patients with non-A, non-B hepatitis. HCV RNA was detectable in 83 of 90(92%)patents. It was detectable in liver tissue in 36 of 40(90%)patients. Thus, it was revealed that mst of patients with non-A, non-B hepatitis were infected with hepatitis C virus. We also detected the change of HCV RNA in patients treated by interferon. When patients were effectively treated, HCV RNA in serum persistently disappeared, in contrast it remained positive when the treatment was resuccessful. Detection of HCV RNA seems to indicate whether the treatment is effective or not. The developrant of detection method of HCV RNA seems to be important for the diagnosis and treatment of type C hepatitis.
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