Background:
Epstein-Barr virus (EBV) is a widespread DNA herpes virus that infects 90% of the healthy population by adulthood. The virus mainly infects B lymphocytes, immortalizing them in culture, but also oropharyngeal epithelial cells and T-cells. EBV has strong epidemiologic links to Infectious Mononucleosis, Burkitt Lymphoma and Nasopharyngeal Carcinoma. There is a weaker link to Hodgkin’s Lymphoma. Post-transplant lymphoprolierative disease (PTLD) is a potentially life-threatening complication after transplant and EBV is a strong risk factor.
Clinical Utility:
Quantitative PCR assay has an important role in monitoring virus reactivation and quantification of EBV, allowing for early treatment intervention and post-treatment observation of PTLD.
Methodology:
This EBV-PCR assay utilizes Nanogen’s MGB Alert Detection Reagent Analyte Specific Reagents for amplification of Epstein-Barr virus. Nucleic acid is extracted from samples and PCR is performed on the ABI 7500 Real-Time PCR System. Following PCR, the MGB probes enable melting curves to be generated and allow for detection of low-level targets as well as variants. EBV amplification and melting curves are observed within the same PCR as the Internal Control amplification and melting curves.
Sensitivity:
270 copies of EBV DNA per milliliter
Specificity:
No cross-reactivity with the following viral species was seen: HSV, HIV, HCV, CMV, VZV and Enterovirus.
Specimen Requirements:
- Whole Blood:
- Adult: 5-10 mL whole blood in purple top (EDTA) tube.
- Peds: 2-5 mL whole blood in purple top (EDTA) tube.
- Pediatric Minimum is 200µL.
- Ship cold, but not frozen.
- Plasma:
- Adult: 5-10 mL whole blood in purple top (EDTA) tube, spin and freeze plasma within 4 hours of collection.
- Peds: 2 – 5 mL whole blood in purple top (EDTA) tube, spin and freeze plasma within 4 hours of collection.
- Pediatric Minimum is 200µL of plasma.
- Ship to laboratory frozen
- CSF:
- Adult: 1-2mL in sterile container.
- Peds: 0.5-1mL in sterile container.
- Pediatric Minimum is 200µL.
- Ship to laboratory cold.
A REQUISITION FORM MUST ACCOMPANY ALL SAMPLES. Please include detailed clinical information.
Test Performed (Days):
Monday, Tuesday, Wednesday, Friday
Turn Around Time:
1-3 days
Shipment Sensitivity Requirements:
Keep whole blood and CSF cold and ship on cold packs. Package and ship plasma on dry ice. Please contact Client Services at (855) 535-1522 for shipping kits and instructions. Ship via overnight express, using the FedEx priority overnight label provided.
References:
- Gulley M, Fan H, Elmore S. 2006. Validation of Roche LightCycler Epstein-Barr Virus Quantification Reagents in a Clinical Laboratory Setting. Journal of Molecular Diagnostics 8(5): 589-597.
- Hayden R, Hokanson K, Pounds S, et al. 2008. Multicenter Comparison of Different Real-Time PCR Assays for Quantitative Detection of Epstein-Barr Virus. Journal of Clinical Microbiology 46(1): 157-163.
- Meerbach A, Wutzler P, Häfer R, et al. 2008. Monitoring of Epstein-Barr Virus Load After Hematopoietic Stem Cell Transplantation for Early Intervention in Post-Transplant Lymphoproliferative Disease. Journal of Medical Virology 80(1): 441-454.
Additional Info: