EBV Conference 2008 Guangzhou - Baylor College of Medicine

206 (RegID: 1293)
Jaap Middeldorp
Institution: VU University medical center, Dept.Pathology
e-mail: j.middeldorp@vumc.nl
RAPID TEST FOR DIAGNOSIS OF MONONUCLEOSIS AND EBV CARRIERSHIP BASED ON
EBV-SPECIFIC PEPTIDES AND MONOCLONAL ANTIBODIES DETECTING IGM-VCA- AND
IGG-VCA/EBNA1/EAD
JM Middeldorp, S Stevens, SAMW Verkuijlen, D Schol and R van Herwijnen
Posterabstract:
Purpose: Acute and active EBV infection is accompanied by a variety of “non-specific” symptoms
resulting from associated inflammatory responses. Similar symptoms are observed in other acute and
chronic diseases requiring specific laboratory testing. Diagnosis of EBV infection demands the use of
multiple tests for the detection of IgM and IgG antibodies to various EBV antigens, which complicates
direct test options (bed side or doctor’s office).
Approach: Using IgM- and IgG-specific monoclonal antibodies and combinations defined biotinylated
synthetic peptides or purified recombinant proteins representing immuno-dominant epitopes of EBV VCA,
EBNA1 and EAd complexes linked to nitrocellulose membranes (capture) and 10-20nm gold particles
(detection signal), we were able to construct simple membrane-based “rapid tests” for the specific
detection of EBV-reactive IgM-VCA or IgG-VCA/-EBNA1/-EAd in human sera with simple handling and
10-15 min. reaction time.
Results: More than 50 sera from mononucleosis (IM) patients were correctly diagnosed by a positive
IgM-VCA and negative EBNA1-IgG reaction. Follow-up samples showed appropriate IgM/IgG-VCA
dynamics, correlating well with IgM-/IgG-ELISA values used as standard. EAd-IgG was temporarily
found in IM patients but not in healthy EBV carriers. Sera from 25 patients with various acute non-EBV
viral infections and autoimmune diseases gave negative EBV-IgM results. RF-positive sera gave no
non-specific results. Sera from healthy EBV carriers were identified by a positive reaction for
IgG-VCA/EBNA1, with a negative response for EAd-IgG. Photometric scanning of the marker bands in
situ, including the positive control, allowed for semi-quantification.
Conclusion: This new approach allows complex EBV diagnostic serology using multiple distinct
antigen-antibody markers to be used in an accurate single rapid test format, permitting direct application
in the doctor’s office.
EBV Conference 2008 Guangzhou
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