Epstein Barr Virus (EBV) RNA Detection by in-situ Hybridization
EBV virus, EB virus, EBER-1
Specimen Collection Criteria
- Collect: Paraffin-embedded tissue. A paraffin block must be submitted. (Slides or paraffin shavings are not acceptable.) Submit formalin-fixed, paraffin-embedded block with corresponding H&E slide. Tissue should be well fixed and well processed. RNA will be assessed for quality during the process of hybridization. If it is deemed unacceptable, testing will be cancelled with client notification.
- Specimen must be accompanied by a completed requisition and must contain the patient name, date of birth, collection date, ordering physician, and source of specimen.
Physician Office/Draw Specimen Preparation
Maintain paraffin-embedded tissue or slides at room temperature (20-25°C or 68-77°F) until transport.
Preparation for Courier Transport
Transport: Paraffin-embedded tissue, at room temperature (20-26°C or 68-78.8°F).
- Tissue in fixatives other than 10% formalin or zinc formalin.
- Decalcified tissue.
- Improper labeling, inadequate information.
Inpatient Specimen Preparation
Please refer to Surgical Specimen, Routine in the Beaumont Laboratory Test Directory
Maintain specimens at room temperature (20-26°C or 68-78.8°F) until testing.
Specimen Stability for Testing:
Room Temperature (20-26°C or 68-78.8°F): Indefinitely
Refrigerated (2-8°C or 36-46°F): Unacceptable
Frozen (-20°C/-4°F or below): Unacceptable
Specimen Storage in Department Prior to Disposal:
Room Temperature (20-26°C or 68-78.8°F): 7 days
Royal Oak Anatomic Pathology – Advanced Diagnostics Laboratory.
Monday – Friday.
Results available in 4 business days.
An interpretive report will be provided.
EBV messenger RNA in infected cells
EBV messenger RNA in infected cells can be detected by a hybridization reaction with oligonucleotide probes (EBER). The test utilizes horseradish peroxidase (HRP)-conjugated detection chemistry to localize fluorescein-labeled oligonucleotide probes. Following the hybridization of fluorescein-labeled probes to target sequences in the tissue, a stringent wash solution containing a blocking agent is used to remove excess bound probe and to block nonspecific binding sites on the tissue that may otherwise react with the detection reagents. The fluorescein label is amplified via a mouse anti-fluorescein isothiocyanate (FITC) linker antibody. This can be visualized with HRP conjugated anti mouse antibody and a colorimetric reaction or the enzyme conjugate with its substrate DAB (3, 3’-diaminobenzidine). This reaction results in the deposition of an insoluble brown product at the site of hybridization.
A pathologist’s interpretation of results will be added to the surgical report.
Latent Epstein-Barr virus (EBV) infection of human tissues results in the expression of EBV RNA (EBER) in infected cells, which can be detected by in-situ hybridization assays using oligonucleotide probes in paraffin sections.
Anatomic Pathology – RO
Advanced Diagnostics Lab – RO
Microtainer® and Vacutainer® are registered trademarks of Becton, Dickinson and Company.
UroVysion® is a registered trademark of Abbott Laboratories. ThinPrep® is a registered trademark of Hologic, Incorporated.