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PubMed1991

Cytomegalovirus testing: antibody determinations and virus cultures with recommendations for use.

Human cytomegalovirus is a ubiquitous virus with the property of becoming latent in the immunocompetent host. It can also cause devastating infections in the immunocompromised patient. Several different assays are available to detect either cytomegalovirus antibodies or cytomegalovirus itself, five of which are used at the University of Wisconsin Hospitals and Clinics and the Wisconsin State Laboratory of Hygiene. The latex agglutination (LA) test is used to detect antibody positivity in transplant donors, transplant recipients, and others. The complement fixation (CF) test is used to detect seroconversion or reactivation of infection in the CMV positive host. The immunofluorescent antibody-IgM (IFA-IgM) test is used to diagnose infection in congenitally or neonatally infected infants. The "routine" CMV culture is the "gold standard" for the detection of the cytomegalovirus in patient secretions. The shell vial assay, a rapid method of detecting virus, is most useful when contemplating therapy for active CMV infection. The advantages and limitations, as well as the timing and appropriateness of each test, are reviewed.

Antibodies, Viral↗
PubMed1985

Comparison of six methods for the detection of antibody to cytomegalovirus.

Five commercial assays were compared to a standardized complement fixation (CF) test for the detection of antibody to cytomegalovirus. Two hundred and thirty serum specimens were analyzed. In addition, nine pairs of acute- and convalescent-phase sera were tested by two of the commercial assays. The assays were compared as to sensitivity, specificity, and positive and negative predictive value, as well as incidence of false-positive and -negative results. Samples which did not agree in all the assays were retested and tested with an indirect fluorescent-antibody assay. Of 228 specimens, 103 (45.2%) were positive by CF. Of the 230 samples, 2 (0.9%) were inconclusive by CF and readable in the other assays. Of the 230 specimens, 97 (42.2%) were positive by an enzyme immunoassay (EIA; Litton Bionetics), 100 (43.5%) were positive by a second EIA (Abbott Laboratories), 104 (45.2%) were positive by a third EIA (M. A. Bioproducts). One hundred and eight (47.0%) were positive by indirect hemagglutination (IHA; Cetus Corporation), and 110 (47.8%) were positive by latex agglutination (LA; Hynson, Westcott and Dunning). Sensitivity and specificity were similar with all the assays (93 to 100%). The greater numbers of positive results by IHA and LA were confirmed by repeat CF testing at less than 1:8 dilution, and by indirect fluorescent-antibody assay. Acute- and convalescent-phase serum pairs showed a significant rise in antibody titer when tested by anticomplement immunofluorescence, IHA, and LA. There was good agreement among the assays, with LA having the highest sensitivity.

Antibodies, Viral↗
PubMed2002

Humoral immune responses following experimental infection of goats with Mycoplasma capricolum subsp. capripneumoniae.

Goats housed in microbiologically secure facilities were experimentally endobronchially infected with Mycoplasma capricolum subsp. capripneumoniae (Mccp), causal agent of contagious caprine pleuropneumonia (CCPP). The animals were monitored over an 8-week period post-infection (p.i.). Elevated temperatures were observed 2-7 days p.i., reaching a maximum of 41.5 degrees C in one animal (1884). By 8 weeks p.i. the infection was successfully cleared, with no Mccp being recovered from the lungs, serum or nasal passages. Mccp was not isolated from serum throughout the experiment, either directly by culture or indirectly via polymerase chain reaction (PCR). Humoral immune responses against Mccp capsular polysaccharide (CPS) were generally poor when measured by ELISA. CPS antigen was present in the serum of all infected animals early in the infection (day 14 p.i.), although in one animal (1855) CPS antigen persisted throughout. This was the only animal to exhibit a serious cough (day 5-19 p.i.). Successful diagnosis of CCPP was achieved using two different types of latex agglutination test (CPS antibody and CPS antigen detection test), immunoblotting and a blocking ELISA, although the latter lacked sensitivity until later in the infection (35-40 days p.i.). Only a single animal (1855) was detected positive using the current complement fixation test (CFT). Strong immune responses to protein antigens were detected by IgG and IgM immunoblotting from the first time point at day 14 p.i. IgM immunodominant bands of 220, 85, 62 and 40kDa were observed in the 3 infected animals and from CFT-positive CCPP field sera. Band intensity gradually diminished throughout the experiment. IgG immunodominant bands of 108, 70, 62, 44, 40 and 23kDa were shared between experimentally-infected and field sera, with band intensity either remaining unchanged or increasing from day 14 p.i. These bands were not present using pre-infection sera. Of the diagnostic tests used, only the CPS antibody detection latex agglutination test and IgG immunoblotting gave positive diagnoses throughout the entire period post-infection (days 14-53 p.i.).

Animals↗
PubMed1979

Circulating immune complexes in mixed connective tissue disease.

Circulating immune complexes were measured by three assays (monoclonal rheumatoid factor radioimmunoassay, C1q-binding assay, Raji cell radioassay) in sera from patients with mixed connective tissue disease. Evidence for circulating immune complexes was found by at least one method in 94% of sera. The highest frequency of positive results was noted by the Raji cell radioassay (88%). However, a significant number of patients also had immune complexes measured by the C1q-binding assay (50%) and monoclonal rheumatoid factor radioimmunoassay (45%). Further, the levels of immune complexes are measured by the Raji cell radioassay, and to a lesser extent the C1q-binding assay, correlated with disease activity and represent a potentially useful clinical tool. The pattern of reactivity in mixed connective tissue disease sera differed from rheumatoid arthritis, scleroderma, and polymyositis. This suggests that a spectrum of immune complexes is found in these rheumatic diseases and supports the concept that mixed connective tissue disease is distinct from these diseases.

Antibodies, Antinuclear↗