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dc.contributor.authorWilliams, David
dc.contributor.authorMenegola, David
dc.contributor.authorMoody, K.
dc.contributor.authorYang, L.
dc.contributor.authorLevy, A.
dc.contributor.authorBriese, T.
dc.contributor.authorTokarz, R.
dc.contributor.authorHarnett, G.
dc.contributor.authorMacKenzie, John
dc.contributor.authorSmith, D.
dc.contributor.authorLipkin, W.
dc.contributor.editorM. Cooley and S. Tristram
dc.identifier.citationWilliams, David and Menegola, David and Moody, Kate and Yang, Lee and Levy, Avram and Briese, Thomas and Tokarz, Rafal and Harnett, Gerald and MacKenzie, John and Smith, David and Lipkin, W. 2011. A MassTag PCR Assay for the Syndromic Detection of Pathogens that can cause Neurological Disease, in M. Cooley and S. Tristram (ed), Australian Society for Microbiology Annual Scientific Meeting 2011, Jul 4-8 2011, pp. 1-149. Hobart, TAS: Australian Society for Microbiology.

MassTag PCR is a novel technology for the rapid, sensitive and simultaneous detection of multiple gene sequences. This technique utilises a library of molecular tags, each unique in its molecular weight. MassTags are conjugated to oligonucleotide primers using a UV-cleavable linker that enables separation of primer and tag. Different primers are labelled each with a different molecular weight tag and are used to amplify target nucleic acids in a multiplex RT-PCR. After removing unincorporated primers, tags are released by UV irradiation and analysed using a single quadrupole liquid chromatography mass spectrometer (LC/MS). Thus, amplification of the gene target produces a unique dual signal in LC/MS analysis that allows its identification. Neurological disease represents a diagnostic challenge, with over 100 microbial agents associated with infection of the central nervous system (CNS). We have developed a panel of multiplex MassTag PCR assays to specifically detect >30 microbial agents that cause CNS infections, relevant to an Australasian diagnostic setting. Three assays were developed using optimised PCR chemistry and thermocycling conditions to detect: (i) common microbial causes of disease (11-plex); (ii) arthropod-borne and zoonotic infections (14-plex); and (iii) rare or uncommon causes (11-plex). The performance of these assays was evaluated in a clinical diagnostic laboratory using CSF specimens and sensitivity estimates for each target were established using synthetic nucleic acids. Our findings indicate that MassTag PCR offers an inexpensive and sensitive diagnostic platform suitable for high-throughput testing.

dc.publisherAustralian Society for Microbiology
dc.titleA MassTag PCR Assay for the Syndromic Detection of Pathogens that can cause Neurological Disease
dc.typeConference Paper
dcterms.source.titleAustralian Society for Microbiology Annual Scientific Meeting 2011 Final Program and Abstract Book
dcterms.source.seriesAustralian Society for Microbiology Annual Scientific Meeting 2011 Final Program and Abstract Book
dcterms.source.conferenceAustralian Society for Microbiology Annual Scientific Meeting 2011
dcterms.source.conference-start-dateJul 4 2011
dcterms.source.conferencelocationHobart, Australia
curtin.departmentSchool of Biomedical Sciences
curtin.accessStatusFulltext not available

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