Next generation decision making
Detecting the threat: novel (rapid) diagnostics. The CIs have extensive experience with development of high-throughput microbial screening and diagnosis (12 - 15): Automated MT-PCR methods have been developed in collaboration with AusDiagnostics for pathogen/R gene quantitation (including mRNA expression) to monitor progress and potential risk markers (1.1,1.2). First developed for ‘avian’ H5N1 influenza (NHMRC #402870 Iredell/Dwyer/Gilbert/Stanley) and widely used in the H1N1/09 pandemic, we have also optimised MT-PCR for bacteria and yeasts (15). We will develop predictive markers for commercial use. Complementary platforms can be tested: time-gated luminescence microscopy (high sensitivity and specificity cf fluorescence microscopy; automated stage and image-recognition software) - developed with a commercial partner, Olympus (ARC #LP0775916 Iredell); rolling-circle PCR and multiplex PCR-reverse line blot [Gilbert], all of which have been either pioneered, developed or optimised extensively in the CI laboratories (13, 14).
12. #Stanley KK, Szewczuk E. Multiplexed tandem PCR: gene profiling from small amounts of RNA using SYBR Green detection. Nucleic Acids Res 2005;33(20):e180.
13. *Steain MC, Dwyer DE, Hurt AC, et al. Detection of influenza A H1N1 and H3N2 mutations conferring resistance to oseltamivir directly on clinical specimens using rolling circle amplification. Antiviral Research 2009; in press.
14. *Kong F, Gilbert GL. Multiplex PCR-based reverse line blot hybridization assay (mPCR/RLB)a practical epidemiological and diagnostic tool. Nat Protoc 2006;1(6):2668-80.
15. *Lau A, Sorrell TC, Chen S, Stanley K, Iredell J, Halliday C. Multiplex tandem PCR: a novel platform for rapid detection and identification of fungal pathogens from blood culture specimens. J Clin Microbiol 2008;46(9):3021-7.