| Project/Area Number |
63570310
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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, Faculty of Medicine, Assistant, 医学部, 助手 (90182691)
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| Project Period (FY) |
1988 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1989: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1988: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | hepatitis B virus / HBV DNA / polymerase chain reaction / HBeAg / Ab / direct sequencing / chronic liver disease / HBs抗原 / HBs抗原サブタイプ / 慢性肝炎 / ポリメラーゼ・チェイン・リアクション法 / B型肝炎ウイルス / 合成DNA / HBVDNA / 超高感度検出法 / HBe抗体 / 肝疾患 |
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| Research Abstract |
Hepatitis B virus is a major etiologic agent of various liver diseases. It is estimated that there are about 2 million virus carriers in Japan. So far, detection of hepatitis B virus DNA (HBV DNA) is performed using Southern blot or spot test method, but the limit of sensitivity by these methods is about 0.1-1.0 picogram (equivalent to 10^4-10^5 viral particles). We established a method to detect HBV DNA at the level of one virion using polymerase chain reaction (PCR) method which can exponentially amplify nucleic acids. Using this method, we detected HBV DNA in sera from HBsAg positive patients with chronic liver diseases. We found HBV DNA in all the sera from HBeAg positive patients. We also found it in the majority of patients with HBeAb positive patients who were supposed to have no HBV DNA in their serum so far. We also revealed that HBeAb positive patients who have normal liver function tests for long period (up to 7 years) have no HBV DNA in serum. Thus these patients were supposed to have cleared off hepatitis B virus from their serum. In addition,we analyzed the PCR product by direct sequencing method and showed analysis of HBV DNA sequence could be easily performed by these methods. With this technique we confirmed the report that the HBsAg subtype d/y and r/w was defined by the amino acid of the 122nd and 160th codon of HBsAg.
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