Enzyme-linked immunosorbent assay for measurement of cytomegalovirus glycoprotein B antibody in serum.
Measurement of antibody to cytomegalovirus (CMV) glycoprotein B (gB) is valuable in the assessment of the antibody response to infection and to gB-containing vaccines. For this purpose, an enzyme-linked immunosorbent assay (ELISA) with a recombinant CMV gB molecule as the antigen was evaluated. Sera from 168 anti-CMV IgG-positive and 100 seronegative subjects were used to evaluate the anti-gB antibody assay. A cutoff optical density (OD) that would distinguish gB antibody-positive from -negative sera was established. Titers of antibody to gB determined by endpoint dilution were compared with those calculated using regression analysis. The run-to-run and interoperator reproducibilities of results were measured. The mean OD + 5 standard deviations from 50 anti-CMV IgG antibody-negative sera (0.2472) was used as the cutoff between anti-gB antibody-positive and -negative results. All sera from 100 anti-CMV IgG-seronegative subjects were negative for antibody to gB. All but 1 of 168 sera from seropositive subjects were positive for antibody to gB. Observed antibody levels based on titration to the endpoint were very similar to results calculated using linear regression. The run-to-run consistency of endpoints was excellent, with 38 runs from one operator and 48 runs from another all giving results within 1 dilution of the mean value for each of three anti-CMV IgG antibody-positive serum pools. The geometric mean titer of antibody to gB for 99 sera from seropositive blood donors was 1/10,937. This ELISA gives accurate and reproducible results for the relative quantity of anti-CMV gB IgG in serum over a wide range of antibody levels.
Hackett
Daniel J
DJ
Department of Pediatrics, University of Alabama at Birmingham, Birmingham, AL 35233, USA.
Zhang
Changpin
C
Stefanescu
Carla
C
Pass
Robert F
RF
D000328:Adult; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral; D019411:Clinical Laboratory Techniques / Q000379:methods*; D003586:Cytomegalovirus Infections / Q000175:diagnosis* / Q000276:immunology; D022482:Cytomegalovirus Vaccines / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D011994:Recombinant Proteins; D015203:Reproducibility of Results; D044967:Serum / Q000276:immunology*; D014759:Viral Envelope Proteins; D055815:Young Adult
D000914:Antibodies, Viral; D000956:Antigens, Viral; D022482:Cytomegalovirus Vaccines; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C040495:glycoprotein B, Simplexvirus