EPSTEIN-BARR VIRUS (EBV) QUANTITATIVE DETECTION
Label Name: EBV QNT DET
Lab Discipline: Molecular Diagnostics
Subdiscipline: Molecular Diagnostics
Institution:  Duke University Health System 
EAP ID:  LAB1373 
Last Review:  3/16/2017 2:50:35 PM
Container & Volume
  Age Group   Container   Volume  
  0  - 18 Years LAVENDER TOP TUBE 1  ML
Label Reminders
  Be sure to include patient's name, history #, date and time of collection, and collector's initials.
Collection Notes
  All:
  • Bone Marrow: One lavender-top EDTA tube (minimum of 1 ml) is required for testing. Forward unprocessed bone marrow promptly at ambient temperatures. THE SPECIMEN CANNOT BE FROZEN. GREEN-TOP (HEPARIN) TUBES ARE NOT ACCEPTABLE FOR TESTING.

    Peripheral Blood: One lavender-top EDTA tube (minimum of 3 mls) is required for testing. Forward unprocessed peripheral blood promptly to the laboratory at ambient temperatures. THE SPECIMEN CANNOT BE FROZEN. GREEN-TOP (HEPARIN) TUBES ARE NOT ACCEPTABLE FOR TESTING.
 
Transport
  Deliver peripheral blood and bone marrow specimens to lab at ambient temperature. If there is a delay of more than 24 hours in delivery, refrigerate the sample. DO NOT FREEZE.
Turn Around Time -  Routine: 5 Days   Stat: N/A
Reference Values
EBV QNT DET
NEGATIVE
Methodology
  This assay uses the Qiagen EBV PCR (ASR) reagents and the ABI PRISM 7500 Sequence Detection System (Applied Biosystems) to detect and quantify nucleic acid sequences specific to EBV genomic DNA. Total DNA is first extracted from whole blood or bone marrow. Real time quantitative PCR is then performed on an aliquot of this DNA. In a single reaction mixture, two PCR primer/ TaqMan probe set combinations are used. The first specifically targets and amplifies a 97 bp conserved sequence in the EBV EBNA-1 gene. The second specifically targets a spiked internal control (IC) which is designed to prevent false negative results due to inefficient extraction or inhibition of PCR amplification. The EBV viral copy number is calculated by interpolation to a standard curve consisting of serially diluted EBV DNA standards. The lower detection limit of this assay is 1 copy of the EBV genome per ul of blood or bone marrow. The linear range for this assay is 25 to 25000 copies of the EBV genome per ul of blood or bone marrow.

This test was developed and its performance characteristics determined by the DUHS Clinical Molecular Diagnostics Laboratory. It has not been cleared or approved by the U.S. Food and Drug Administration. This test is used for clinical purposes. It should not be regarded as investigational or for research. This laboratory is certified under the Clinical Laboratory Improvement Amendments of 1988 ("CLIA") as qualified to perform high complexity clinical testing.
   
   
Clinical Significance and Interpretive Data
    BACKGROUND:

Epstein-Barr virus (EBV), one of the eight human herpes viruses (HHV4), is a ubiquitous double-stranded DNA virus. The virus is usually transmitted by salivary contact and most individuals become infected during childhood. In children, primary infection is usually asymptomatic or accompanied by mild nonspecific symptoms. After resolution of a primary infection, the virus establishes a lifelong latent infection within B-lymphocytes and the epithelial cells of the oral cavity and nasopharynx.

A number of EBV associated neoplastic processes have been described. In immunocompetent individuals these include Burkitt lymphoma, Hodgkin lymphoma and nasopharyngeal carcinoma. In individuals who are immunocompromised due to bone marrow or solid organ transplantations, EBV can be re-activated to cause post-transplant lymphoproliferative disorder (PTLD). A number of EBV associated neoplasms in AIDS patients have also been described including smooth muscle tumors, AIDS-related Hodgkin lymphoma, and AIDS-related non-Hodgkin lymphomas (including CNS lymphoma, plasmablastic lymphoma, and primary effusions lymphoma).

INTERPRETATION:

• Numerical results are reported as EBV copies per µl of whole blood or bone marrow.

• When possible, EBV copy number should be correlated with the clinical status of the patient and with prior EBV copy number.

• Since EBV infection is highly prevalent, an individual may have a low copy number of EBV DNA present in his or her peripheral blood but not have an EBV associated neoplasm or disease. Please contact the Clinical Molecular Diagnostics Laboratory for additional questions or for help interpretting your results.

REFERENCES:

1. Humar et al. (2006) American Society of Transplantation Recommendations for Screening, Monitoring and Reporting Infectious Complications in Immunosuppression Trials in Recipients of Organ Transplantation. American Journal of Transplantation 6: 262-274.

2. Gulley and Tang (2008) Laboratory Assays for Epstein-Barr Virus-Related Disease Journal of Molecular diagnostics 10(4): 279-292.

3. Kimura et al (2008) Measuring Epstein-Barr virus (EBV) load: the significance and application of each EBV-associated disease. Reviews of Medical Virology 18(5): 305-319.
   
Indications
    EBV testing is RECOMMENDED for the following patients:

• Patients who are at high risk of developing post-transplant lymphoproliferative disorder (PTLD). This includes; patients who have received a solid organ or bone marrow transplant who are heavily T-cell immunosupressed (eg. having received anti-thymocyte antibodies as part of their treatment), transplant patients who were EBV naďve prior to transplant, and individuals who have received an HLA mismatched transplant. A rising EBV titer in these patients is suggestive of re-activation or new infection by EBV, and could indicate the development of PTLD. A rising EBV titer alone is not diagnostic of PTLD. Additional studies are required to make a definitive diagnosis. These include histologic analysis, flow cytometry and/or IGH clonality studies.

• Patients who have been diagnosed with PTLD. By following the titer of EBV in the peripheral blood of a patient, a physician can monitor a patient’s response to therapy or progression of disease.

EBV testing CAN BE CONSIDERED in the following patients:

• Patients with nasopharyngeal carcinomas. In these patients, EBV copy number can be used for prognosis; a high EBV copy number is suggestive of advanced stage disease.

• Patients with EBV-related lymphomas. These include AIDS-related Hodgkin lymphoma, AIDS-related non-Hodgkin lymphomas (including CNS lymphoma, plasmablastic lymphoma, and primary effusions lymphoma), and a variety of Hodgkin and non-Hodgkin lymphomas in immune competent patients (including mixed cellularity Hodgkin lymphoma, Burkitt Lymphoma, Pyothorax-associated lymphoma, Lymphomatoid granulomatosis, Primary effusion lymphoma, Angioimmunoblastic T-cell lymphoma, Extranodal NK/T lymphoma of the nasal type, and others). For some of these patients, a onetime measurement of peripheral blood EBV titers can have prognostic value. The value of monitoring EBV levels as an indicator of response to therapy in these patients has not been proven and remains controversial.

• AIDS patients with the clinical suspicion of primary CNS lymphoma. In these patients EBV levels are elevated in the cerebrospinal fluid. A high EBV copy number in the CSF of a patient with a clinically suspicious radiologic lesion can provide a working diagnosis of CSF lymphoma (in conjunction with other studies including flow cytometry, IGH clonality and cytologic evaluation of CSF WBCs) if biopsy of the lesion is not possible.
   
Contraindications
    The appropriate test for patients with the clinical suspicion of infection mononucleosis is EBV serology. In these patients a hetophile antibody test ('mono spot test') is sufficient for diagnosis.
   
Limitations
    .The lower detection limit of this assay is 1 copy of the EBV genome per µl of whole blood or bone marrow.

• This test detects copy numbers of EBV DNA in whole blood and bone marrow. It does not distinguish between latent EBV infection and active EBV infection.

• An elevated EBV copy number is not diagnostic of any one disease process.

• This test is subject to interference by various factors such as DNA quality, presence of PCR inhibitor, mutation and polymorphism of EBNA-1 gene sequence. Such factors could produce a false negative result.
   
Test Synonyms
  Synonym(s): EBV
Synonym(s): nasopharyngeal carcinoma
Synonym(s): PTLD
Molecular Diagnostics Laboratory
(MDX)

Medical Director:
 Michael Datto, M.D., Ph.D.
 Phone: 919-684-6965
 Email: michael.datto@duke.edu
Lab Director:
 Catherine Rehder Ph.D, FACMG
 Phone: 919-613-8434
 Email: catherine.rehder@duke.edu
Lab Director:
 Siby Sebastian Ph.D., DABMG
 Phone: 919-613-8432
 Email: siby.s@duke.edu

Address: 
 Wadsworth Bldg, Cytogenetics, Rm 0220
 2351 Erwin Rd
 Durham,  NC  27705
 Phone: 919-684-2698
 FAX: 919-668-5424

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