NEW MATERIAL:A specific antigen protein of hepatitis of non-A non-B type containing an amino sequence shown by the formula or part thereof and a DNA fragment coding the protein.
USE: An antigen protein is effective for detecting an antibody against hepatitis of non-A non-B type and a DNA fragment is useful as a probe for detecting a gene of specific antigen related to hepatitis of non-A non-B type by nucleic acid hybridization and a reagent for genetic diagnosis such as primer of genetic amplification.
PREPARATION: RNA of hepatitis virus of non-A non-B type is extracted from plasma, serum or liver tissue of patient of hepatitis of non-A non-B type. cDNA is obtained from the prepared RNA fraction by random primer method, inserted into bacteriophage λ gt11 and Escherichia coli Y1,090 strain, etc., is infected with the bacteriophage. Plaque produced by culture is transferred to a nitrocellulose membrane and treated with serum retaining specific antibody of hepatitis of non-A non-B type to detect an antibody reaction positive clone. Escherichia coli is infected with the clone, amplified and the objective DNA fragment is isolated. The DNA fragment is manifested to give an antigen protein.
HATANAKA TADASHI
NANBA TOSHIHIKO
YAMADA KYOKO
TSUJI MASAO
ARIMA TERUMASA