Real-Time Quantitative EBV PCR

Epstein Barr Virus may be detected in a wide variety of clinical specimens including saliva, plasma, serum, oral swabs, and other bodily fluids. This virus has long been known to be associated with Burkitt ’s lymphoma, a malignancy that is decidedly present and of great clinical significance in sub-Saharan Africa. HIV increases the risk of developing Burkitt's Lymphoma as well as complicating its treatment.

The extraction of viral DNA from the sample can be done by different procedures using a variety of different techniques. Qiagen chemistry using both a high-throughput and single column procedure, is an effective and efficient method that is currently being used by our lab.

After DNA is extracted, Real-Time Taqman PCR technology utilizing fluorescent labeled probes is used to amplify, detect and quantify a variety of EBV viral DNA in the sample. Primers and probes specific to the BALF5 region of the EBV genome have been designed and tested for sensitivity and specificity at the University of Washington Molecular Diagnostic Lab in Seattle, Washington, USA.  An internal control is spiked into each PCR reaction to ensure that no non-specific inhibition of the reaction has taken place. A negative result is accepted only if the internal control amplification is detected.