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Epstein Barr Virus (EBV) RNA Detection by in-situ Hybridization

EBV virus, EB virus, EBER-1

Department

Advanced Diagnostics

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). 

Rejection Criteria

• 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.

In-Lab Processing

Maintain specimens at room temperature (20-26°C or 68-78.8°F) until testing.

Storage

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

Laboratory

Royal Oak Anatomic Pathology – Advanced Diagnostics Laboratory.

Performed

Monday – Friday.
Results available in 4 business days.

Reference Range

An interpretive report will be provided.

Test Methodology

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. 

Interpretation

A pathologist’s interpretation of results will be added to the surgical report.

Clinical Utility

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. 

CPT Codes

88365

88364x2.

Contacts

Last Updated

9/29/2022