Rapid Detection of Human Cytomegalovirus and Herpesvirus Hominis IgM Antibody by the Immunoperoxidase Technique

Gerna, Giuseppe; Chambers, Robert

doi:10.1159/000148901

Cited by 17 publications

(5 citation statements)

References 11 publications

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“…It may be assumed that most of these patients had a reactivation of a previous CMV infection or a reinfection with CMV. Other investigators (1,6,16) have also observed that a proportion of patients with evidence or reactivation or reinfection with CMV did produce IgM antibody.…”

Section: Discussionmentioning

confidence: 89%

Direct enzyme-linked immunosorbent assay that uses peroxidase-labeled antigen for determination of immunoglobulin M antibody to cytomegalovirus

Loon¹,

Heessen²,

Logt³

et al. 1981

J Clin Microbiol

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A direct enzyme-linked immunosorbent assay was developed for the measurement of immunoglobulin M (IgM) antibody to cytomegalovirus (CMV). Wells of microtiter plates were coated with anti-human IgM. Each patient's serum was added at a dilution of 1:100, and IgM from the serum was allowed to react with anti-human IgM. The amount of CMV-specific IgM antibody bound was determined by measuring the intensity of color change after the addition of peroxidaselabeled CMV antigen and substrate. Nuclei of infected cells served as an antigen source. CMV IgM could be detected only in IgM fractions of sera from patients with a recent CMV infection. Rheumatoid factor did not cause false-positive results. No cross-reactions were observed when paired sera from 22 patients with herpes simplex or varicella and single sera from 12 patients with suspected infectious mononucleosis were tested by the direct enzyme-linked immunosorbent assay. Each of 17 patients with a seroconversion for CMV antibody showed CMVspecific IgM antibody. In six of these patients the antibody was detected in the initial serum. The direct enzyme-linked immunosorbent assay for CMV IgM is a specific and sensitive test for the diagnosis of recent CMV infections and possesses distinct advantages over indirect tests. 18 to 72 years of age. Four patients were children 3, 4, 9, and 15 years of age, none of whom had been congenitally infected with CMV. In addition, sera were obtained from 34 patients clinically suspected of having herpes simplex or varicella-zoster infection or infectious mononucleosis. Furthermore, 36 sera with 416

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Section: Discussionmentioning

confidence: 89%

Direct enzyme-linked immunosorbent assay that uses peroxidase-labeled antigen for determination of immunoglobulin M antibody to cytomegalovirus

Loon¹,

Heessen²,

Logt³

et al. 1981

J Clin Microbiol

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“…In the group of five reactivated CMV infections, CMV elimination was always associated with detection of EA-Ab at different titers. In the three groups of subjects (healthy people, leukemic children, and pregnant women) tested for presence of CMV EA-Ab, the prevalence of this antibody was 6 of 31 leukemic children, 4 of 28 pregnant women, and 3 of 17 healthy people.…”

Section: Resultsmentioning

confidence: 98%

Immunoglobulin G to virus-specific early antigens in congenital, primary, and reactivated human cytomegalovirus infections

Gerna¹,

Cereda²,

Cattaneo³

et al. 1978

Infect Immun

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Immunoglobulin G antibody to human cytomegalovirus (CMV)-specific early antigens (EA-Ab) was determined by the immunoperoxidase antibody technique in several cases of congenital, primary, and reactivated CMV infections. Mothers of congenitally infected infants and a group of leukemic children and pregnant women were also studied. In 11 cases of congenital infection, CMV EA-Ab was always associated with CMV excretion whether immunoglobulin M antibody was present or not. Nine mothers of congenitally infected infants had CMV EA-Ab for several months after delivery, but association with CMV elimination was not established when urine and/or saliva were tested for virus isolation. In all nine cases of primary CMV infection, CMV EA-Ab was present, and in five its detection was associated with CMV isolation. In one case, disappearance of EA-Ab occurred when virus excretion ceased. In five cases of reactivated CMV infections, a consistent association between CMV EA-Ab and virus isolation was found. Six of 31 leukemia children had CMV EA-Ab, and virus was isolated from 3 of these. Four of 28 pregnant women showed EA-Ab in their serum, but tests for isolation were not done. These data suggest that CMV EA-Ab is not a marker of a current primary CMV infection, as previously reported, but a marker of an active CMV replication which can take place in primary as well as in congenital and reactivated CMV infections.

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“…In developing the test, the type of CMV antigen used for coating of polystyrene beads was of greatest importance. Antigens prepared from CMV-infected cell suspensions, the most commonly used antigen in CF tests and in previously reported RIA [Knez et al, 19761 and ELISA tests [Gerna et al, 1977[Gerna et al, , 1978Cappel et al, 1978;Ziegelmaier et al, 1978; P h a and Pejtsik, 19791 produced high background values, unsatisfactory specific binding, and titration curves with inverted slope in IgG assays. A possible explanation for this may be CMV-induced Fc-binding, which is supported by the facts that the control antigen gave markedly less cpm than cell-associated CMV antigen when used in titrations of negative reference sera in IgG assays and that the inverted slope was not seen with the control antigen.…”

Section: Dlscusslonmentioning

confidence: 99%

“…Detection of cytomegalovirus (CMV)-specific antibodies by enzyme-linked immunosorbent assay (ELISA) and solid-phase radioimmunoassay (RIA) has been described by several authors [Knez et al, 1976;Gerna et al, 1977Gerna et al, , 1978Cappel et al, 1978;Ziegelmaier [Sarov et al, 19801 of these studies, antigens prepared from infected cell suspensions were used. Among possible pitfalls in CMV serology are false-positives due t o CMV-induced IgG-(Fc) receptors [Furukawa et al, 1975;Betts et al, 1976;Keller et al, 1976;Rahman et al, 1976;Griffiths et al, 19781, anticell antibodies [The and Langenhuysen, 1972;Anderson and Andersen, 19781 and rheumatoid factors [Gispen et al, 1975;Knez et al, 1976;Meurman et al, 1977;Cremer et al, 1978;Krishna et al, 19801, The present report describes a radioimmunoassay in which extracellular CMV harvested by ultracentrifugation was adsorbed on polystyrene beads and antibodies that reacted with the virus antigen were assayed by iodinated anti-human IgG, IgM, and IgA.…”

Section: Introductionmentioning

confidence: 99%

Solid‐Phase Radioimmunoassay of Serum IgG, IgM, and IgA Antibodies to Cytomegalovirus

Torfason

Källander

Halonen

1981

Journal of Medical Virology

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A radioimmunoassay (RIA) using polystyrene beads as the solid phase for cytomegalovirus (CMV) antigen and iodinated immunosorbent purified anti-human IgG, IgM, and IgA as indicator antibodies was developed for the detection of immunoglobulin class-specific antibodies to CMV. An antigen prepared from extracellular virus was essential for reliable results, and a preparation ultracentrifuged and sonicated twice was better than a crude antigen. The optimal antigen gave low cpm values with a negative reference serum, resulting in cpm ratios of 10 or higher between early convalescent phase serum and negative reference serum. Of six patients with an increase in CMV CF titres, all six had an increase in RIA IgG titres, four had an increase in IgA titres, and all had IgM antibodies. The IgG titres were high, up to 1/64,000. In a group of 17 infants negative in CMV CF test, 14 had CMV IgG antibodies in RIA test, indicating mainly low levels of maternal antibodies. In six of seven patients with CMV isolations from urine specimens, an increase in IgG or IgA titres or the presence of IgM antibodies was found, and only one of these patients had an increase in CMV CF titre. The specificity of the developed CMV RIA test was further demonstrated by detecting no significant increase in RIA titres in serum specimens of patients with primary herpes simplex infection, chickenpox, herpes zoster, or infectious mononucleosis.

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Rapid Detection of Human Cytomegalovirus and Herpesvirus Hominis IgM Antibody by the Immunoperoxidase Technique

Cited by 17 publications

References 11 publications

Direct enzyme-linked immunosorbent assay that uses peroxidase-labeled antigen for determination of immunoglobulin M antibody to cytomegalovirus

Direct enzyme-linked immunosorbent assay that uses peroxidase-labeled antigen for determination of immunoglobulin M antibody to cytomegalovirus

Immunoglobulin G to virus-specific early antigens in congenital, primary, and reactivated human cytomegalovirus infections

Solid‐Phase Radioimmunoassay of Serum IgG, IgM, and IgA Antibodies to Cytomegalovirus

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