[Preliminary use of recombinant glycoprotein 52kd as an antigen in the diagnosis of human cytomegalovirus infection].
Interest in the human cytomegalovirus (HCMV) mainly derives from its associations with congenital malformations, mental retardation, and severe or fatal infections in immunosuppressed individuals such as transplant patients, tumor and AIDS patients. It is evidenced that there has been a need for a rapid and sensitive methods to detect an ongoing acute infection. The recent studies showed that high titers of antibody to the glycoprotein 52kd are present in sera of patients undergoing acute HCMV infection. However, purification of individual glycoprotein from HCMV-infected cells is a daunting prospect. HCMV glycoprotein 52 kd expressed via recombinant DNA techniques are a promising approach to solve this problem. In order to evaluate the diagnostic value of the recombinant glycoprotein 52 kd antigenic code region for HCMV infection, we have used the polymerase chain reaction (PCR) and recombinant DNA techniques to construct successfully the high-level expression plasmid pHCMV containing the HCMV GP-52 kd antigenic code region, with the predicted protein at levels up to 20% in total bacterial protein. The expressed protein was purified from SDS-PAGE, used as an antigen in Western-blot, and reacted with 12 cases of the positive sera, 4 cases of the negative sera, following by reaction with HRP-labelled horse IgG antibody against human. The results indicated that the approach we are using to detect antibody to HCMV acute infection are as sensitive as general serological methods such as ELISA, with the advantages of easy preparation of antigen with high quantity, and clinical practicability.
Wu
J
J
Department of Pediatrics, Shanghai Cancer Institute.
Tang
Z
Z
Chen
J
J
Gu
J
J
Wan
D
D
Qu
S
S
Li
H
H
D000914:Antibodies, Viral / Q000032:analysis; D000957:Antigens, Viral, Tumor; D003587:Cytomegalovirus / Q000276:immunology*; D003586:Cytomegalovirus Infections / Q000175:diagnosis*; D006801:Humans; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins / Q000276:immunology
D000914:Antibodies, Viral; D000957:Antigens, Viral, Tumor; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C040612:glycoprotein 52 antigen, Mouse mammary tumor virus