Epstein-Barr Virus (EBV), Molecular Detection, Quantitative PCR, Blood
NY State Approved Indicates the status of NY State approval and if the test is orderable for NY State clients.
A prospective and diagnostic marker for the development of posttransplant lymphoproliferative disorders (PTLD), especially in Epstein-Barr virus (EBV)-seronegative organ transplant recipients who receive anti-lymphocyte globulin for induction immunosuppression and OKT-3 treatment for early rejection
LightCycler Polymerase Chain Reaction (PCR)
(PCR is utilized pursuant to a license agreement with Roche Molecular Systems Inc.)
Reporting Name A shorter/abbreviated version of the Published Name for a test; an abbreviated test name
Epstein-Barr Virus PCR, Quant, B
EBV (Epstein-Barr Virus)
EBV PCR Quant
Epstein Barr Virus
Epstein-Barr virus PCR
Post Transplant Lymphoproliferative Disorder (PTLD)
EBV PCR Quant
Epstein Barr Virus
Epstein-Barr virus PCR
Post Transplant Lymphoproliferative Disorder (PTLD)
Specimen Type Describes the specimen type needed for testing
Whole Blood EDTA
Specimen Required Defines the optimal specimen. This field describes the type of specimen required to perform the test and the preferred volume to complete testing. The volume allows automated processing, fastest throughput and, when indicated, repeat or reflex testing.
Container/Tube: Lavender top (EDTA)
Specimen Volume: 5 mL
Forms: If not ordering electronically, submit a Microbiology Request Form (Supply T244) with the specimen.
Specimen Minimum Volume Defines the amount of specimen required to perform an assay once, including instrument and container dead space. Submitting the minimum specimen volume makes it impossible to repeat the test or perform confirmatory or perform reflex testing. In some situations, a minimum specimen volume may result in a QNS (quantity not sufficient) result, requiring a second specimen to be collected.
Mild OK; Gross reject
Specimen Stability Information Provides a description of the temperatures required to transport a specimen to the laboratory. Alternate acceptable temperature(s) are also included.
|Whole Blood EDTA||Refrigerated||7 days|
Clinical Information Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test
Primary infection with Epstein-Barr virus (EBV), a DNA-containing virus classified among the family Herpesviridae, may cause infectious mononucleosis resulting in a benign lymphoproliferative condition characterized by fever, fatigue, sore throat, and lymphadenopathy. Infection occurs early in life, and by 10 years of age, 70% to 90% of children have been infected with this virus. Usually, infection in children is asymptomatic or mild and may be associated with minor illnesses such as upper respiratory tract infection, pharyngitis, tonsillitis, bronchitis, and otitis media. The target cell for EBV infection is the B lymphocyte. Immunocompromised patients, lacking antibody to EBV, are at risk for acute EBV infection that may cause lymphoproliferative disorders in organ transplant recipients (posttransplant lymphoproliferative disorders [PTLD]) and AIDS-related lymphoma.(1) The incidence of PTLD ranges from 1% for renal transplant recipients, to as high as 9% for heart/lung transplants, and 12% for pancreas transplant patients.
EBV DNA can be detected in the blood of patients with this viral infection; however, quantitative evaluation of EBV DNA has been shown to correlate highly with the subsequent (3-4 months) development of PTLD in susceptible patients.(2) Organ transplant recipients seronegative (risk for primary EBV infection) for EBV (frequently children) who receive anti-lymphocyte globulin for induction immunosuppression and OKT-3 treatment for early rejection are at highest risk for developing PTLD compared to immunologically normal individuals with prior infection with this virus.(4,5)
Reference Values Describes reference intervals and additional information for interpretation of test results. May include intervals based on age and sex when appropriate. Intervals are Mayo-derived, unless otherwise designated. If an interpretive report is provided, the reference value field will state this.
Increasing copy levels of Epstein-Barr virus (EBV) DNA in serial specimens may indicate possible posttransplant lymphoproliferative disorders (PTLD).
Positive results are quantitated in copies/mL.
Reportable range is 2,000 to 200,000,000 copies/mL. Specimens with results <5,000 copies EBV DNA/mL include a disclaimer that states: "Results may not be reproducible due to low copy number." Blood specimens of normal blood donors for EBV infection usually have low or undetectable levels of viral DNA.
Cautions Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances
Serial determination of blood specimens from patients may be necessary to monitor increasing (risk of development of posttransplant lymphoproliferative disorders [PTLD]) or decreasing (treatment efficacy) levels of Epstein-Barr virus (EBV) DNA.
The reference is range is typically "negative" for this assay, and 20 whole blood samples from "normal" asymptomatic donors were negative for the presence of EBV DNA by.this assay. However, viremia or viral shedding may occasionally be detected in asymptomatic individuals. Therefore, this assay is only to be used for patients with a clinical history and symptoms consistent with EBV infection, and must be interpreted in the context of the clinical picture. This test should not be used to screen asymptomatic patients.
Twenty-five specimens from patients with prior infection with Epstein-Barr virus (EBV) were assayed for EBV-DNA by conventional PCR (gel and Southern blot). Quantitative levels of EBV-DNA were determined using a competitive target (plasmid containing target EBV-DNA). The results were recorded as EBV copies/mL in the following range: 0-50; 51-500; 501-5000; 5001-50,000. This preliminary conventional PCR assay has been formatted to a real-time LightCycler PCR which allows the determination of specific numeric copy levels of EBV-DNA.
All 25 samples, indicated above, were tested by LightCycler PCR for EBV-DNA. Specific copy levels obtained for all 25 samples correlated with the ranges obtained by conventional PCR.
To supplement the above data,50 whole blood samples submitted to the Mayo Clinic Virology Lab were tested by a Mayo laboratory-developed EBV PCR assay and the Roche EBV ASR PCR reagent. After resolution of 1 discrepant sample (noted to be very low copy number), the sensitivity and specificity were 100% and 86%, respectively.
Analytical Sensitivity/Limit of Detection (LoD):
The lower limit of detection (LoD) for this assay is 1 target/ microliter in whole blood.
No PCR signal was obtained from extracts of 40 bacterial and viral isolates that could cause similar symptoms including herpes simplex virus (HSV) 1 and 2, cytomegalovirus (CMV), varicella-zoster virus(VZV) and human herpesvirus (HHV) HHV6, HHV 7 and HHV 8. Additionally, 20 whole blood samples from "normal" asymptomatic donors were negative for the presence of EBV DNA.
Inter-assay precision was 100% and intra-assay precision was 100%. The Standard Deviation did not exceed 0.1.
Reportable (Linear) Range/Range of Quantification:
This LC PCR assay is linear from 2,000 copies/mL to 200,000,000 copies/mL. The lowest and highest calibrators (expected LLoQ and ULoQ) fall within the linear range. Results will be reported as <2,000 copies/mL, a numeric value between 2,000 copies/mL and 200,000,000 copies/mL or >200,000,000 copies/mL.
Clinical Reference Provides recommendations for further in-depth reading of a clinical nature
1. Paya CV, Fung JJ, Nalesnik MA, et al: Epstein-Barr virus-induced posttransplant lymphoproliferative disorders. ASTS/ASTP EBV-PTLD Task Force and the Mayo Clinic Organized International Consensus Development Meeting. Transplantation 1999;68(10):1517-1525
2. Kenagy DN, Schlesinger K, Weck JH, et al: Epstein-Barr virus DNA in peripheral blood leukocytes of patients with posttransplant lymphoproliferative disease. Transplantation 1995;60(6):547-554
3. Green MJ, Bueno D, Rowe G, et al: Predictive negative value of persistent low Epstein-Barr virus viral load after intestinal transplantation in children. Transplantation 2000;70(4):593-596
4. Kogan DL, Burroughs M, Emre S, et al: Prospective longitudinal analysis of quantitative Epstein-Barr virus polymerase chain reaction in pediatric liver transplant recipients. Transplantation 1999;67(7):1068-1070
5. Green M, Cacciarelli TV, Mazariegos GV, et al: Serial measurement of Epstein-Barr viral load in peripheral blood in lymphoproliferative disease. Transplantation 1998;66(12):1641-1644
6. Lau AH, Soltys K, Sindhi RK, et al: Chronic high Epstein-Barr viral load carriage in pediatric small bowel transplant recipients. Pediatr Transplant Jan 20
Method Description Describes how the test is performed and provides a method-specific reference
Quantitative Epstein-Barr virus (EBV) DNA determinations are obtained by LightCycler PCR.
Viral DNA is extracted by the MagNA Pure automated instrument (Roche Applied Science) from whole blood. LightCycler PCR primers and probes have been designed to detect target EBV DNA. Amplified product is detected by a fluorescence assay (fluorescence resonance energy transfer [FRET]). Quantitative results are obtained by incorporating known copy levels (5 standards) of EBV DNA (plasmid) into the assay together with the sample. LightCycler instrument software calculates the level of EBV DNA (copy/mL) in the sample from a standard curve of the quantitative levels of the reference plasmid EBV DNA.(4,5) (Espy MJ, Patel R, Paya C, Smith TF: Quantification of Epstein-Barr virus viral load in transplant patients by LightCycler PCR. ASM Press 101(st) General Meeting [Abstract]. 2001;May 20-24)
Day(s) and Time(s) Test Performed Outlines the days and times the test is performed. This field reflects the day and time the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time required before the test is performed. Some tests are listed as continuously performed, which means assays are performed several times during the day.
Monday through Saturday; Varies
Analytic Time Defines the amount of time it takes the laboratory to setup and perform the test. This is defined in number of days. The shortest interval of time expressed is "same day/1 day," which means the results may be available the same day that the sample is received in the testing laboratory. One day means results are available 1 day after the sample is received in the laboratory.
Same day/1 day
Maximum Laboratory Time Defines the maximum time from specimen receipt at Mayo Medical Laboratories until the release of the test result
Specimen Retention Time Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded
Performing Laboratory Location The location of the laboratory that performs the test
Test Classification Provides information regarding the medical device classification for laboratory test kits and reagents. Tests may be classified as cleared or approved by the US Food and Drug Administration (FDA) and used per manufacturer's instructions, or as products that do not undergo full FDA review and approval, and are then labeled as an Analyte Specific Reagent (ASR), Investigation Use Only (IUO) product, or a Research Use Only (RUO) product.
This test was developed using an analyte specific reagent. Its performance characteristics were determined by Mayo Clinic in a manner consistent with CLIA requirements. This test has not been cleared or approved by the U.S. Food and Drug Administration.
CPT Code Information Provides guidance in determining the appropriate Current Procedural Terminology (CPT) code(s) information for each test or profile. The listed CPT codes reflect Mayo Medical Laboratories interpretation of CPT coding requirements. It is the responsibility of each laboratory to determine correct CPT codes to use for billing.
LOINC® Code Information Provides guidance in determining the Logical Observation Identifiers Names and Codes (LOINC) values for the result codes returned for this test or profile.
|Result ID||Reporting Name||LOINC Code|
|82987||Epstein-Barr Virus PCR, Quant, B||In Process|
|26399||Lower 95% Confidence Interval||N/A|
|26400||Upper 95% Confidence Interval||N/A|