An approach to identify the etiological agent of Crohn's disease by molecular cloning technique
| Project/Area Number |
04670406
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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 | Akita University |
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Principal Investigator |
ISHIOKA Tomonori (1994) Akita University Medical Department Assistant, 医学部, 助手 (10146772)
千葉 満郎 (1992-1993) 秋田大学, 医学部, 助教授 (90150250)
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| Co-Investigator(Kenkyū-buntansha) |
IIZUKA Masahiro Akita University Medical Department Assistant, 医学部, 助手 (00241654)
NAKAGOMI Osamu Akita University Medical Department Professor, 医学部, 教授 (70143047)
CHIBA Mitsuro Akita University Medical Department Assistant Professor, 医学部, 助教授 (90150250)
石岡 知憲 秋田大学, 医学部, 助手 (10146772)
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| Project Period (FY) |
1992 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1994: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1993: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1992: ¥800,000 (Direct Cost: ¥800,000)
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| Keywords | Crohn's disease / Molecular biology / Gene / Immunoscreening / PCR (polymerase chain reaction) / Measles virus / Inflammatory bowel disease / Crohn病、(クローン病) / Inflammatory bowel disease(IBD) / Crohn病,クローン病 / 炎症性腸疾患 L(Inflammatory bowel disease:IBD) |
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| Research Abstract |
Crohn's disease is a chronic inflammatory bowel disease with unknown etiology. This study was undertaken to seek by molecular techniques the possible etiological agents of Crohn's disease. A cDNA library was constructed from the affected colonic tissue of a Crohn's disease patient. A total of 120*10^6 of lambda phage clones were screened with the serum taken from another patient with Crohn's disease. The immunoscreening selected 20 phage clones containing 150-1000 bp-cDNA inserts. Nucleotide sequencing revealed, however, that 12 of 20 such positive inserts exhibited no homology with any of viral or bacterial gene sequences deposited in the GenBank and European Molecular Biology Laboratory (EMBL) databases.
Because measles virus has recently been reported as an etiological agent of Crohn's disease, we attempted to amplify portions of the viral genome by using the PCR (polymerase chain reaction) with primers specific to the genes encoding nucleocapsid, matrix, fusion protein and hemagglutinin.Single distinct bands with and expected molecular size were observed in all specimens by PCR with primers for the nucleocapsid gene, while no fragment was amplified by any of the other primer pairs. Sequencing these fragments revealed, however, no homology with the nucleocapsid gene of measles virus or any other known sequences deposited in the GenBank and EMBL databases.
From these results, we conclude that no infectious agent, measles virus in particular, is directly involved in the pathogenesis of Crohn's disease.
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Report
(4 results)
Research Products
(3 results)
