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04.355 - CPMV RNA Mimics
Institute for Animal Health (Pirbright, UK)
John Innes Centre (Norfolk, UK)
Ultrasensitive methods for detection of specific RNAs, such as qPCR, are in common use for diagnostics, screening and population monitoring. RNA is a notoriously fragile molecule and it is therefore desirable to include a standard RNA in samples in order to prevent false negative results. The standard RNA must be highly stable under normal conditions, but readily released and amplified by standard sample processing protocols.
Scientists at the Institute of Animal Health, Pirbright, and the John Innes Centre, Norwich, have devised a method of generating stable, non-infectious plant virus particles that encapsulate a chimæric RNA, containing any desired control sequence, or multiple nested control sequences. These control RNA sequences are engineered into the RNA-2 hemigenome of Cowpea Mosaic Virus (CPMV). The virus particles, and the RNA molecules within them, are stable in the presence of biological material at 37°C for over a month, in solution at up to 60°C and 50%DMSO and are highly resistant to RNAse treatment. The encapsulated RNA is readily released by standard lysis protocols (e.g. RNAzol) and amplifies efficiently.
Development of a novel recombinant encapsidated RNA particle:Evaluation as an internal control for diagnostic RT-PCR. Donald P King, Nick Montague, Katja Ebert, Scott M Reid, Juliet P Dukes, Lysann Schädlich, Graham J Belsham, George P Lomonossoff. J Virol Methods 2007. In press (available online August 2007).
Contact: Dr Martin Stocks
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