Assay of measles virus IgM and IgG class antibodies by use of peroxidase-labelled viral antigens

Abstract

Purified measles virions, nucleocapsid protein and crude lysate of measles infected cells were labelled with horseradish peroxidase, and used for the detection of IgM and IgG antibodies to measles virus by direct enzyme immunoassay. The assays consisted of three layers: anti-human IgM or IgG immunoglobulins on solid-phase, test serum specimen and enzyme labelled viral antigen. For the expression of the results, a standard curve was included in each test and the O.D. values were changed to arbitrary antibody units. Specificity and sensitivity of the assays were compared with indirect EIAs. The specificity studies included a collection of serum specimens containing either rheumatoid factor, antinuclear antibodies or IgM antibody specific for other viruses. The assays proved both reliable and simple to perform and sensitivity was slightly higher than that for indirect EIAs. However, specificity was dependent on the purity of the viral antigens. When crude infected cell lysate antigen was used, some nonspecific results were obtained, particularly with serum specimens containing antinuclear antibodies. When virion or nucleocapsid protein were used, no nonspecific reactions were obtained.

Detection of IgM Antibodies against Canine Distemper Virus in Dog and Mink Sera Employing Enzyme-Linked Immunosorbent Assay (ELISA)

“…The ELISA techniques have been developed for the detection of antibodies against the closely related measles virus, and these techniques, which we essentially based on measurements of single serum samples, are currently used for serodiagnosis of present or recent measles virus infections in humans. 15,18,21,22 The principle of these ELISA techniques, in contrast to other CDV ELISA techniques, 10,14,25 is based on an immunological separation of from the test sample before the reaction with viral antigen. This capture technique eliminates competing antibodies and minimizes nonspecific reactions due to rheuma factors in human sera.…”

Design and evaluation of an electrochemical immunosensor for measles serodiagnosis using measles-specific Immunoglobulin G antibodies

“…It also further confirms that the oxidation of TMB only occurs in the presence of H 2 O 2 which initializes the enzymatic activity towards TMB. The monitoring of the presence of measles-specific 11PAb has been accomplished using reported spectrophotometric detection via indirect immunoassays protocols [19,20,44]. Spectrophotometric detection systems are laboratory confined and therefore research is ongoing into electrochemical detection systems which offer onsite monitoring.…”

Characteristics of different solid-phase immunoassay formats for the measurement of BK virus immunoglobulin M in sera of patients on renal dialysis or with kidney allografts

“…RF interference has therefore necessitated the removal of IgG antibody by treatment with either staphylococcal protein A, aggregated IgG, or IgG-coated latex particles prior to testing (4,11,29,32). Additional approaches for improving specificity have included the use of purified viral antigen to eliminate false-positive IgM results obtained with crude antigens (11,28,29) antigen to allow identification of false-positive reactions in antibody capture assays (11).…”

“…Serological studies have shown that pregnant women may have a primary or reactivated BKV infection (6), and this has raised the question of in utero papovavirus infection. Polyomavirus is transmitted transplacentally in mice (19), but evidence for transplacental transmission of polyomavirus in humans is contradictory (6,14,26,28,30) and there have been no confirmed reports of in utero infection in humans.…”