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

在分离、净化和扩增过程中，用于诊断或筛查目的的粗制样品的RNA扩增可能在许多阶段失败。这种失败会产生假阴性结果，这在筛选关键样本时是不可接受的，例如在动物或人类爆发流行病毒疾病期间。因此，即使不是必要的，也是非常必要的，在这个过程中尽早包括控制RNA，这样一个积极的控制信号就可以验证从样品获取到最终扩增和检测的每一个步骤。然而，RNA是一种脆弱的分子，没有保护的RNA标准很快就会降解。我们已经证明，通过将RNA包裹在植物病毒颗粒(N-capRNA)中可以防止这种降解，而且，这种颗粒可以作为高效的管内控制。我们现在需要制定方法，为广泛的目标疾病迅速制定这种控制措施。

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.