EPIDEMIC NON-A NON-B HEPATITIS VIRUS ANTIGEN PEPTIDE AND NUCLEIC ACID FRAGMENT CODING THE SAME

摘要

PURPOSE:To obtain a peptide characteristic to non-A non-B hepatitis virus by using an immunoscreening technique adopting a new molecular genetic means different from conventional immunoserological means and applying the screening technique to a bile sample of a crab-eating macaque infected with non-A non-B hepatitis virus. CONSTITUTION:Bile of a crab-eating macaque infected with non-A non-B hepatitis virus is diluted with physiological saline water and tissue-pieces, etc., are removed by low-speed centrifugation. The solution is placed on a physiological saline water containing 20% sucrose and subjected to ultracentrifugation to recover the virus particles as a precipitate. The particles are treated with guanidine thiocyanate and the extracted RNA is used as a template for the synthesis of cDNA. The cDNA is inserted into lambdagt11 vector to prepare a cDNA library. E.coli is infected with lambda-phage, scattered on a plate for the culture of bacteria, cultured at 42 deg.C for several hours and a replica is produced therefrom. The replica is treated with a blocking liquid, washed with tris-buffer physiological saline water (TBS) and subjected to immunoscreening.