Molecular Detection by Quantitative PCR

Quantitative PCR detection of Epstein-Barr Virus (EBV) DNA.

Plasma, whole blood, CSF, tissue biopsies. Required volume of plasma and whole blood - 2 mL (minimum 1mL); CSF 600 uL (minimum 300 uL). Minimum volume will not allow repeat testing of samples with equivocal results.

For plasma samples, collect blood in plasma preparation tube (PPT) or K2EDTA tubes (plastic; lavender or pink top). Do not use heparin as an anti-coagulant. Centrifuge tube at room temperature at 1,100 RCF for 10-15 min. Aliquot plasma into separate tube. Collect CSF in a dry tube subject to classic conditions for carrying out lumbar punctures.

Do not freeze whole blood, store and ship refrigerated within 48h of sample collection.. Plasma and CSF specimens may be stored and shipped refrigerated to the NML within 48h of sample collection or must be stored and shipped frozen. Fresh biopsies should be stored frozen at all times and shipped on dry ice.

Shipping of specimens shall be done by a TDG certified individual in accordance with TDG regulations. For additional information regarding classification of specimens for the purposes of shipping, consult either Part 2 Appendix 3 of the TDG Regulations or section 3.6.2 of the IATA Dangerous Goods Regulations as applicable.

Specimens are accepted from immunosuppressed patients with suspected EBV reactivation or lymphoproliferative disease, from immunocompetent patients with severe or chronic EBV-related infections, or patients with suspected CNS infection.
**Please note specimens are not accepted for the management of EBV infection in transplant patients.

Completed "Viral STI, Polyoma and Herpesvirus Testing" requisition form including sender lab name, address and telephone number, patient identifier, date of birth or age, sex, specimen reference #, type of specimen, date collected, test requested, reason for testing and any other relevant clinical information.

Contact lab in advance when sending tissue biopsies.

DNA is extracted from specimens and tested by RT-PCR (1) for the EBNA-1 gene. The results are reported as copy number per mL of specimen.

14 calendars days. STAT samples may be considered under special circumstances. Please contact lab to inquire about shorter turn around time.

1. Stevens, S., Verkuijlen, J., Middledorp, J. 2004. Quantitative detection of Epstein-Barr Virus DNA in clinical specimens by Rapid Real-Time PCR targeting a highly conserved region of EBNA-1. Methods in Molecular Biology 292:15-25.