Vol. 8, No. 2
JOURNAL OP CLINICAL MICROBIOLOGY, Aug. 1978, p. 153-159 0095-1137/78/0008-0153$02.00/0 Copyright@) 1978 American Society for Microbiology
Printed in U.S.A.
Analysis of Antibody Assay Methods and Classes of Viral Antibodies in Serodiagnosis of Cytomegalovirus Infection NATALIE E. CREMER, MARJORIE HOFFMAN, AND EDWIN H. LENNETTE Viral and Rickettsial Disease Laboratory, Califorma State Department of Health, Berkeley, Califonua 94704
Received for publication 5 May 1978 Forty-nine serum pairs with antibody to cytomegalovirus (CMV) were evaluated for rises in antibody titer (- fourfold) by indirect hemagglutination (IHA) and complement fixation (CF), using a freeze-thaw antigen (FT) and a glycine extract antigen (GE). In this sample CF-FT detected more rises in antibody titer than did CF-GE. IHA detected the least number. The apparent reason for stationary antibody titers with CF-GE and IHA was the presence of high antibody titers in the first serum specimen. Separation of immunoglobulin classes of 20 serum pairs by sucrose gradient centrifugation indicated that these antibodies with IHA were of the immunoglobulin M (IgM) clas and those with CF-GE were of the IgG class. By separation of immunoglobulin classes, rises in IgG CMV antibody titers were seen with IHA, rises not observed in the whole serum because of high IgM antibody titers in the first serum specimen. Absence of rises in antibody titers with CF-FT was due in part to too early sampling of the second serum specimen (21 21 66.7 90.5 71.5 of viral infection were tested for CMV antibody by CF >10/>21 77.6 87.8 Average 67.4 with two CMV antigens and by IHA. The sera were from patients with clinical signs compatible with CMV a Time of serum collection after onset of symptoms. b infection; 10 of the 49 patients had, in addition, organ Percentage of serum pairs showing a rise in titer transplants, and five had recently undergone heart (-fourfold). surgery. Forty-six of the 49 paired sera were from adults whose average age was 34.5 years, with a range TABLE 2. GM titer of the first serum samples in of 19 to 77 years. Three paired sera were from children relation to a rise in titer between the first and of 1, 4, and 12 years of age, none of whom had been second serum samples congenitally infected with CMV. Twenty-three paired GM titer of first serum samples sera were selected for further study of the immunoglobulin class of CMV antibodies. Selection was made With rise in anti- With no rise in Test on the basis of a fourfold or greater rise in antibody body titer in sec- antibody titer in one or more of titer between the serum pairs by the ond sample second sample tests and by presence of antibody in the fi-st specimen 28 + 5b 776 * 3 CF-GE by one or more of the tests. 8±2 CF-FT 9i3 RF. Sera were checked for rheumatoid factor (RF) 47±6 IHA 512±5c by using Hyland Laboratories' latex globulin reagent in the slide test. Positive specimens were further evalaSera collected
JOURNAL OP CLINICAL MICROBIOLOGY, Aug. 1978, p. 153-159 0095-1137/78/0008-0153$02.00/0 Copyright@) 1978 American Society for Microbiology
Printed in U.S.A.
Analysis of Antibody Assay Methods and Classes of Viral Antibodies in Serodiagnosis of Cytomegalovirus Infection NATALIE E. CREMER, MARJORIE HOFFMAN, AND EDWIN H. LENNETTE Viral and Rickettsial Disease Laboratory, Califorma State Department of Health, Berkeley, Califonua 94704
Received for publication 5 May 1978 Forty-nine serum pairs with antibody to cytomegalovirus (CMV) were evaluated for rises in antibody titer (- fourfold) by indirect hemagglutination (IHA) and complement fixation (CF), using a freeze-thaw antigen (FT) and a glycine extract antigen (GE). In this sample CF-FT detected more rises in antibody titer than did CF-GE. IHA detected the least number. The apparent reason for stationary antibody titers with CF-GE and IHA was the presence of high antibody titers in the first serum specimen. Separation of immunoglobulin classes of 20 serum pairs by sucrose gradient centrifugation indicated that these antibodies with IHA were of the immunoglobulin M (IgM) clas and those with CF-GE were of the IgG class. By separation of immunoglobulin classes, rises in IgG CMV antibody titers were seen with IHA, rises not observed in the whole serum because of high IgM antibody titers in the first serum specimen. Absence of rises in antibody titers with CF-FT was due in part to too early sampling of the second serum specimen (21 21 66.7 90.5 71.5 of viral infection were tested for CMV antibody by CF >10/>21 77.6 87.8 Average 67.4 with two CMV antigens and by IHA. The sera were from patients with clinical signs compatible with CMV a Time of serum collection after onset of symptoms. b infection; 10 of the 49 patients had, in addition, organ Percentage of serum pairs showing a rise in titer transplants, and five had recently undergone heart (-fourfold). surgery. Forty-six of the 49 paired sera were from adults whose average age was 34.5 years, with a range TABLE 2. GM titer of the first serum samples in of 19 to 77 years. Three paired sera were from children relation to a rise in titer between the first and of 1, 4, and 12 years of age, none of whom had been second serum samples congenitally infected with CMV. Twenty-three paired GM titer of first serum samples sera were selected for further study of the immunoglobulin class of CMV antibodies. Selection was made With rise in anti- With no rise in Test on the basis of a fourfold or greater rise in antibody body titer in sec- antibody titer in one or more of titer between the serum pairs by the ond sample second sample tests and by presence of antibody in the fi-st specimen 28 + 5b 776 * 3 CF-GE by one or more of the tests. 8±2 CF-FT 9i3 RF. Sera were checked for rheumatoid factor (RF) 47±6 IHA 512±5c by using Hyland Laboratories' latex globulin reagent in the slide test. Positive specimens were further evalaSera collected
