What tests should I order if Epstein-Barr Virus (EBV) infection is suspected?

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Last updated: February 17, 2025View editorial policy

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From the Guidelines

Diagnostic Approach for Epstein-Barr Virus (EBV) Infection

EBV infection should be suspected and tested for using a combination of complete blood count (CBC) with differential, liver function tests (LFTs), and EBV-specific serologic tests, including IgM and IgG antibodies to viral capsid antigen (VCA) and Epstein-Barr nuclear antigen (EBNA) 1.

  • Monospot test (heterophile antibody test) may also be performed to aid in diagnosis, although it may yield false-negative results in approximately 10% of patients, especially in children younger than 10 years 1.
  • EBV antibody testing for IgG and IgM to VCA and EBNA is recommended when rapid Monospot or heterophile test results are negative, to differentiate EBV infection from a mononucleosis-like illness caused by other pathogens 1.
  • The presence of VCA IgM (with or without VCA IgG) antibodies in the absence of antibodies to EBNA indicates recent primary infection with EBV, while the presence of EBNA antibodies indicates infection more than 6 weeks from the time of the sample 1.
  • In certain cases, such as in immunocompromised patients or when monitoring disease progression, EBV polymerase chain reaction (PCR) or EBV viral load testing may be necessary to confirm the diagnosis or monitor disease progression 1.

It is essential to interpret these test results in the context of clinical presentation and other laboratory findings to ensure accurate diagnosis and appropriate management of EBV infection.

From the Research

Diagnostic Tests for Epstein-Barr Virus (EBV) Infection

To diagnose EBV infection, several tests can be ordered, including:

  • Immunoglobulin G (IgG) and IgM antibodies to the viral capsid antigen (VCA) 2, 3, 4
  • Heterophile antibodies (HA) 2, 3
  • IgG antibodies to the EBV early antigen-diffuse (EA-D) and nuclear antigen (EBNA-1) 3
  • EBV PCR to detect EBV DNA in serum 5, 6
  • In situ hybridization (ISH) of EBV-encoded RNA (EBER) in biopsy tissues for histopathological lesions 6

Interpretation of EBV Serological Patterns

The interpretation of EBV serological patterns can be challenging due to the multiple possible patterns 3. However, the use of tables to interpret EBV serological patterns based on three or five markers can help to interpret >95% of BioPlex serological results 3.

Choice of Diagnostic Tests

The choice of diagnostic tests may depend on the clinical presentation and the suspected stage of EBV infection. For example:

  • For primary EBV infection, EBV PCR can be used to confirm a positive IgM VCA in the absence of heterophilic antibodies 5
  • For EBV reactivation, the utility of EA titers is disputed, and EBV PCR may be a more reliable method 5
  • For histopathological lesions, ISH of EBER in biopsy tissues remains the gold standard for detecting EBV 6

Comparison of Diagnostic Methods

Different diagnostic methods have been compared in terms of their performance, including:

  • Immunofluorescence assay (IFA) as the reference method 4
  • Enzyme-linked fluorescent assay (ELFA) and immunochromatographic assay (ICA) as suitable methods for the diagnosis and staging of EBV 4
  • Immunoblot assay and enzyme immunoassay (EIA) as less reliable methods for certain EBV antibodies 4

References

Guideline

Guideline Directed Topic Overview

Dr.Oracle Medical Advisory Board & Editors, 2025

Research

Prevalence of primary versus reactivated Epstein-Barr virus infection in patients with VCA IgG-, VCA IgM- and EBNA-1-antibodies and suspected infectious mononucleosis.

Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology, 2007

Research

Real-time Epstein-Barr virus PCR for the diagnosis of primary EBV infections and EBV reactivation.

Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology, 2005

Research

Recent Advances in Diagnostic Approaches for Epstein-Barr Virus.

Pathogens (Basel, Switzerland), 2020

Professional Medical Disclaimer

This information is intended for healthcare professionals. Any medical decision-making should rely on clinical judgment and independently verified information. The content provided herein does not replace professional discretion and should be considered supplementary to established clinical guidelines. Healthcare providers should verify all information against primary literature and current practice standards before application in patient care. Dr.Oracle assumes no liability for clinical decisions based on this content.

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