Evaluation of 12 Commercial Tests for Detection of Epstein-Barr Virus-Specific and Heterophile Antibodies

Abstract

The test panel included sera from patients with primary EBV infection, immunocompromised patients with recent cytomegalovirus infection, healthy persons (blood donors), and EBV-seronegative persons. Among the tests for EBV-specific antibodies the sensitivity was good, with only small differences between the different assays. However, there was a greater variation in specificity, which varied between 100% (Enzygnost) and 86% (Biotest). Tests for detection of heterophile antibodies based on purified or selected antigen (Avitex, Alexon, Clearview IM, and Cards؎OS Mono) were more sensitive than the Paul-Bunnell-Davidsohn and Monosticon tests.The diagnosis of infectious mononucleosis is usually based on typical clinical and hematologic findings and confirmed with a positive test for heterophile antibodies. However, in some cases there is a need for analysis of Epstein-Barr virus (EBV)-specific antibodies, especially when there are atypical symptoms or in the absence of heterophile antibodies. This is often observed with specimens from children, who also may have an unusual EBV antibody pattern. Tests used for the virological diagnosis of mononucleosis should have both high sensitivity and high specificity.In Norway, tests for detection of both EBV-specific and heterophile antibodies are performed at the majority of the microbiological laboratories. Our regular quality assessment system has revealed the need for better control with the use of commercial tests. Also, over the past few years, some newly developed tests have been introduced. Together with nine other microbiological laboratories, the Department of Virology at the National Institute of Public Health (NIPH) in Oslo, Norway, conducted an evaluation of a total of 12 commercial tests for detection of EBV-specific and heterophile antibodies. MATERIALS AND METHODSParticipants in the study. The microbiological departments in the following counties in Norway participated in the study: Aust-and Vest-Agder, Akershus, Bergen, Nordland, Oslo (NIPH, the National Hospital, and Ullevål Hospital), Rogaland, Sogn and Fjordane, and Vestfold.Tests evaluated. Table 1 gives information on the EBV serological tests evaluated, including manufacturers, antigens, test principle, and mode of detection. All enzyme-linked immunosorbent assay (ELISA) tests were based on microwell enzyme immunoassay (EIA). Table 2 gives similar information on tests for heterophile antibodies.Test panel. A test panel consisting of 248 and 241 serum specimens for the EBV antibody and heterophile antibody evaluations, respectively, was selected on the basis of both clinical diagnosis and results of laboratory investigations. Before distribution to the sites, the sera received code numbers. Specimens were assigned to the following groups.(i) Group A. Group A consisted of specimens from patients with recent primary EBV infection. The diagnosis was confirmed by a positive test for heterophile antibodies and an EBV antibody pattern compatible with recent primary infection. A total of 139 and 140 serum specime...

Application of Virus-Specific Immunoglobulin M (IgM), IgG, and IgA Antibody Detection with a Polyantigenic Enzyme-Linked Immunosorbent Assay for Diagnosis of Epstein-Barr Virus Infections in Childhood

“…The Enzygnost EBV ELISA system for the diagnosis of EBV infection is an accurate system for the determination of virus-specific IgG and IgM antibodies (2,3,8,15,19) and has also been shown to detect reactivated EBV infection in nonage-defined populations (4). However, according to our findings, the application of these ELISAs in a pediatric population is characterized by a high rate of indeterminate test results (15 of 66 samples [22.7%]), which points to a relevant imperfection in test performance, because clear-cut interpretation and diagnosis are not possible for these patients.…”

“…Diagnosis of the different stages of EBV infection is based on the determination of EBV-specific immunoglobulin M (IgM), IgG, and IgA antibodies with this assay. The detection of anti-EBV antibodies of the IgM and IgG classes is specific and sensitive for the identification of primary or past EBV infections (2,3,8,9,15,19), and determination of IgA anti-EBV levels in patients with enhanced IgG anti-EBV antibody values (Ͼ650 U/ml) enables chronic or reactivated EBV infections to be diagnosed (4). However, no evaluation of the Enzygnost-based diagnosis of primary, recent, and prolonged or reactivated EBV infections in childhood is available.…”

Serological diagnosis of Epstein-Barr virus infection: Problems and solutions

“…the P3HR-1 or Raj cell lines) [2,23,45,[50][51][52] , whereas EIAs use purified native or recombinant proteins, synthetic peptides or fusion proteins (complete proteins or fragments of the proteins encoded by the EBV genes) [42,48] . The type and preparation of the antigens used are probably responsible for the differences in the sensitivity and specificity of the various assays [42,53,54] . IFA has been used as the reference method, although its sensitivity is the same as or less than that of EIAs [43] , automated versions of which allow a large number of samples to be tested and are commonly used in laboratories with a large routine workload.…”

EBV infection in immunocompetent patient: problems and solutions in serological diagnosis

“…(8,38,43,65,68,89), mentre i test EIA usano proteine native purificate o ricombinanti, proteine di fusione o peptidi sintetici rappresentanti sia proteine complete che frammenti delle proteine codificate dai geni di EBV (31,106). Il tipo e la preparazione degli antigeni utilizzati sono probabilmente responsabili delle differenze riscontrate nei risultati con i vari test (31) sia in termini di sensibilità che di specificità (12,16). Gli ultimi CLIA, inoltre, usando peptidi sintetici con differenti cut off per gli anti-VCA IgM e anti-EBNA-1 IgG possono discriminare meglio lo stadio dell'infezione (29) e, in caso di campioni simultaneamente anti-EBNA-1 IgG, anti-VCA IgG e anti-VCA IgM positivi, possono aiutare nel distinguere tra infezione recente (in fase transitoria) e infezione pregressa o riattivazione (29,73).…”