Vol. 10, No. 2

JOURNAL OF CLINICAL MICROBIOLOGY, Aug. 1979, p. 256-258 0095-1 137/79/08-0256/03$02.00/0

Detection of Adenovirus Antibody by Counterimmunoelectrophoresis MARTIN PETRIC, PETER J. MIDDLETON,* AND SADANAND D. MANKIKAR The Hospital for Sick Children, Toronto, Ontario, Canada

Received for publication 18 May 1979

Counterimmunoelectrophoresis was adapted as a rapid, sensitive assay for detection of adenovirus group-specific antibody. This assay was six times more sensitive than the complement fixation test and was successfully adapted for use with microliter quantities of serum or whole blood. Routine laboratory adenovirus serology is based chiefly on the complement fixation (CF) test. This is a rather uncomplicated group-specific assay (2). Other assays include the neutralization test (7) and hemagglutination inhibition test (3, 5). Both of these tests detect type-specific antibody and are also more sensitive than the CF test (6). A test using tannic acid-treated, adenovirus-coated erythrocytes has been developed, though it has not gained wide acceptance

(1).

2 to 1:64 was determined for 55 adenovirus CIEP-positive sera. Table 2 shows the ratio of CIEP to CF titers. These varied from 0.5 to 64 with the mean CIEP being about six times as high as the overall mean CF titer. From this limited study it may be concluded that the CIEP test was not only substantially more efficient but also more sensitive than the CF test. In this regard it has yet to be rigorously proven that the CIEP test did not give false-positive results. However, the use of twofold CsCl gradient-purified virus (which contained no foreign matter when examined by electron microscopy) as the antigen makes this possibility remote. In our previous publication (4) it appeared that the CIEP test was not significantly affected by impurities of the specimens. No purification

On investigating adenovirus antigen-antibody interactions by counterimmunoelectrophoresis (CIEP) we observed that this system was more sensitive than CF for detection of group-specific antibody. We report here on the comparison of the two systems and on the simplification of of the stools was necessary, since the agarose gel CIEP assay for adenovirus serology. The CIEP was carried out as reported in the was impermeable to the larger particulate mataccompanying manuscript (4). The antigen was ter from the specimen while allowing the antiCsCl gradient-purified adenovirus type 5. The gens and serum proteins to migrate and interact. sera were collected for routine serology from a The feasibility of detecting antibody from a variety of patients admitted to The Hospital for whole-blood specimen by CIEP was therefore Sick Children. Serum samples were stored fro- examined. A total of 40 blood specimens was collected zen at -20°C. The whole-blood samples and associated sera were collected from volunteer from our laboratory, nursing, and resident staffs. staff members. All CIEP slides were read under A portion of each was stored unclotted in a code by two observers. In the event of discord, heparinized tube, while the remainder was alwhich occurred very rarely, the negative value lowed to clot for serum collection. After treatwas accepted. ment at 56°C the sera were tested for adenovirus In the first study (Table 1) a total of 79 sera antibody by CF and CIEP. The whole bloods from hospital patients were tested for adenovi- were tested for adenovirus antibody by CIEP. rus antibody by CIEP and CF over a serial The CIEP assay on whole blood detected 24 dilution range of 1:2 to 1:8. Thirty-seven of these donors with adenovirus antibody. The same aswere positive for adenovirus antibody by CF and say on sera from the same donors detected 27 55 were positive by CIEP. Thus CIEP detected with antibody to adenovirus. The efficiency of 18 additional adenovirus-positive sera that had blood CIEP as compared to serum CIEP for a CF titer of

Detection of adenovirus antibody by counterimmunoelectrophoresis.

Vol. 10, No. 2 JOURNAL OF CLINICAL MICROBIOLOGY, Aug. 1979, p. 256-258 0095-1 137/79/08-0256/03$02.00/0 Detection of Adenovirus Antibody by Counteri...
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