Testing for Adventitious Retroviruses Using PCR-Based Revese Transcriptase Assays

Product enhanced reverse transcriptase (PERT) assays, also termed Amp-RT or polymerase chain reaction-based reverse transcriptase (PBRT) assays (1), are extremely sensitive tests for the detection of reverse transcriptase (RT). They have been reported to be up to 106-fold more sensitive than conventional RT assays for detecting the presence of retroviruses (2,3,4,5). The tests are also the first choice for detection of RT in live viral-vaccines, gene therapy preparations and the screening of animals and patients for Porcine endogenous retrovirus (PoERV) thought to be of concern in xenotransplantation. The assays involve converting an RNA template to cDNA and then amplifying the cDNA using product-specific primers. Since no exogenous RT activity is added to the reaction, cDNA will only be generated if the sample itself contains RT activity.