NUCLEIC ACID FRAGMENT CODING NON-A NON-B TYPE HEPATITIS VIRUS ANTIGEN PEPTIDE AND METHOD FOR UTILIZING SAME FRAGMENT

摘要

PURPOSE:To enable detection of non-A non-B type hepatitis virus in lever structure by cloning a causative virus (gene) of non-A non-B type hepatitis virus. CONSTITUTION:A plasma obtained by adding a precipitating agent such as polyethylene glycol to a human plasma and ultracentrifuging the plasma and concentrated to about 1000 times is treated with guanidium thioacetate and extracted with phenol/chloroform and precipitated with ethanol to purify total nucleic acid in concentrated plasma. Then cDNA synthesized from DNA obtained by decomposing and purifying human- derived DNA containing deoxyribonuclease is inserted into lambdagt11 vector to prepare cDNA library. Then replica obtained by culturing Escherichia coli infected by lambda phage and then culturing the phage infected Escherichia coli using a nitrocellulose filter is reacted with a human or chimpanzee serum at recovery stage or acute stage of non-A non-B type hepatitis and then reacted with enzyme labeled antihuman IgG or IgM and reacted with a substrate solution to color plague and a phage corresponding to the colored plague is selected and subjected to secondary screening to provide the nucleic acid fragment coding non-A non-B type hepatitis virus antigen peptide.