ABSTRACT
The primary and secondary structures of the pp65 phosphoprotein of human cytomegalovirus coded by the UL83 gene were studied by the methods of computer-aided analysis. An immunodominant protein fragment with 3 antigenic determinant was detected. The UL83 fragment coding the selected region was amplified and cloned in bacterial expressing vector. The recombinant protein was obtained and purified. On the basis of ELISA findings it was acknowledged as possible to use the pp65 recombinant protein jointly with pp150 and p52 in the diagnosis of antibodies specific to human cytomegalovirus.
Subject(s)
Cytomegalovirus/metabolism , Phosphoproteins/genetics , Viral Matrix Proteins/genetics , Amino Acid Sequence , Enzyme-Linked Immunosorbent Assay , Epitopes/chemistry , Molecular Sequence Data , Phosphoproteins/chemistry , Phosphoproteins/metabolism , Protein Structure, Secondary , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Sequence Homology, Amino Acid , Viral Matrix Proteins/chemistry , Viral Matrix Proteins/metabolismABSTRACT
Designing of a liquid serum panel for the determination of class G antibodies specific to human cytomegalovirus (HCMV) is under discussion. Sera were selected by ELISA for antibodies to HCMV and by PCR for CMV DNA. The serum panel comprises samples of positive and negative sera with high and low titers. Sera were stabilized by a stabilization solution. The panel shelf life was evaluated by routine methods and by the "accelerated aging" technique. Sera selected for a standard panel containing or not antibodies to HCMV preserve their properties and stability for as long as 1 year at 4 degrees C.
