PSEUDORABIES VIRUS LATENCY - A QUANTITATIVE APPROACH BY POLYMERASE CHAIN-REACTION

Some data dealing with the establishment of a quantitative PCR assay a re presented. The assay is based on the use of an internal standard (m imic) which differs from the target by a deletion of a few base pairs and which is co-amplified with the target DNA. The resulting PCR produ cts are labelled with fluorescent primers and then separated and detec ted by an automated sequencer. A highly specific and sensitive PCR ass ay for the envelope glycoprotein gp50 gene has been developed. This as say is highly reproducible with a detection limit of one copy of PRV D NA. Several mimics were then constructed. As a result we can confirm t hat the strategy that we have chosen is adequate for the quantificatio n of low amounts of virus DNA present in latently infected swine.