N-Cap RNA: protected RNAs for in-tube diagnostic controls and inter-laboratory standards

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Amplification of RNA from crude samples for diagnostic or screening purposes can fail at many stages during the isolation, clean-up and amplification processes. Such failures produce false negative results, which are not acceptable when screening critical samples, for instance during outbreaks of epidemic viral disease in animals or humans. It is therefore highly desirable, if not essential, to include control RNA at the earliest possible step in the process, so that a positive control signal can validate every step from sample acquisition to final amplification and detection. However RNA is a fragile molecule and an unprotected RNA standard is very rapidly degraded. We have shown that this degradation can be prevented by encapsulating the RNA within a plant virus particle (N-cap RNA) and, furthermore, that such particles can act as highly effective in-tube controls. We now need to develop methods for the rapid production of such controls for a wide range of target diseases.