Community Research and Development Information Service - CORDIS

Development of real-time PCR methods to detect and quantify MVV proviral DNA

Real-time PCR tests to detect and quantify MVV EV1 pro-virus DNA and mRNA in blood and tissues were developed. The primers and dual-labeled probes were designed on the gag and pol genes of EV1. Comparison of the relative sensitivities of the gag and pol assays showed that the gag assay was superior. The sensitivity of the gag real-time assay is approximately 3 copies/reaction. The real-time PCR tests can be used to detect and quantify pro-virus DNA or mRNA (after reverse transcription) in the blood and tissues of experimental animals involved in vaccines studies (as in this project) or in pathogenesis studies. The assays could also be used to detect MVV in routine diagnostic settings, though the assays are specifically designed to detect the EV1 strain and have not been assessed for sensitivity and specificity in relation to field strains.

The end users of the information are other scientists working in the field of SRLV control, or working with other lentiviruses, or working with other plasmid DNA vaccines, or in the general area of vaccines and immune responses, or in the general area of lentivirology.

Commercial companies may be interested in the real-time PCR tests, but further work is required to determine if the assays have any value in routine diagnosis. Also, the gag and pol PCR tests were designed specifically to detect MVV EV1 strain for the purposes of the project, and were not designed with the routine diagnosis of SRLV in the field in mind. It is unlikely that the assays will be commercialised without an assessment of their value in the field.

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