RSE/BBSRC Enterprise Fellowship Dr Nicholas Montague

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Reliable positive controls are vital when performing diagnostic testing of potential disease samples. Without robust positive controls there is always the possibility of producing a false negative result. The potential seriousness of such an outcome when testing for a disease such as foot-and-mouth cannot be exaggerated. The highly sensitive method of real-time reverse-transcription PCR (rRT-PCR) is now the routine front-line tool for diagnosis of viruses in clinical samples. Guidelines for the validation and quality control of these PCR-based tests require internal controls to be included within the assay. The current controls used are artificial RNA constructs. However this 'naked' RNA is far from ideal as a control as it is prone to degradation and, more importantly, cannot accurately follow the diagnostic protocol from start to finish. The genetic material of viruses, such as FMDV, is protected within a protein 'shell' and must be extracted prior to PCR testing; naked RNA does not require this. In order for a control to accurately follow this step in the procedure we have developed a novel method of encapsulating RNA within a cowpea mosaic virus (CPMV) viral capsid. These non-pathogenic chimaeric virus particles would be added to the potential disease sample taken directly from the suspect animal and taken through the entire diagnostic process. In addition, diagnostic laboratories worldwide are required to undergo proficiency testing to insure uniformity of results. To this end accurately quantified and identical disease samples must be used at each laboratory. Containment and security concerns prohibit the sending of viral disease samples to these laboratories, however the encapsidated mimics would be ideal for this purpose. It is proposed that the use for proficiency testing will be licensed to the appropriate regulatory bodies. In the future the encapsidated mimics could be trialled for use within for human medical diagnostics.

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