title: A nanoplasmonic label-free surface-enhanced Raman scattering strategy for non-invasive cancer genetic subtyping in patient samples.
abstract: Simple nucleic acid detection methods could facilitate the progress of disease diagnostics for clinical uses. An attractive strategy is label-free surface-enhanced Raman scattering (SERS) due to its capability of providing structural fingerprinting of analytes that are close to or on nanomaterial surfaces. However, current label-free SERS approaches for DNA/RNA biomarker detection are limited to short and synthetic nucleic acid targets and have not been fully realized in clinical samples due to two possible reasons: (i) low target copies in limited patient samples and (ii) poor capability in identifying specific biomarkers from complex samples. To resolve these limitations and enable label-free SERS for clinical applications, we herein present a novel strategy based on multiplex reverse transcription-recombinase polymerase amplification (RT-RPA) to enrich multiple RNA biomarkers, followed by label-free SERS with multivariate statistical analysis to directly detect, identify and distinguish between these long amplicons (∼200 bp). As a proof-of-concept clinical demonstration, we employed this strategy for non-invasive subtyping of prostate cancer (PCa). In a training cohort of 43 patient urinary samples, we achieved 93.0% specificity, 95.3% sensitivity, and 94.2% accuracy. We believe that our proposed assay could pave the way for simple and direct label-free SERS detection of multiple long nucleic acid sequences in patient samples, and thus facilitate rapid cancer molecular subtyping for personalized therapies.
authors.affiliation: Centre for Personalised Nanomedicine, Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Brisbane, QLD 4072, Australia. j.wee@uq.edu.au yuling.wang@mq.edu.au m.trau@uq.edu.au.
authors.forename: Jing
authors.identifier: http://orcid.org/0000-0001-6080-7998
authors.initials: J
authors.lastname: Wang
authors.affiliation: Centre for Personalised Nanomedicine, Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Brisbane, QLD 4072, Australia. j.wee@uq.edu.au yuling.wang@mq.edu.au m.trau@uq.edu.au.
authors.forename: Kevin M
authors.identifier: http://orcid.org/0000-0002-2652-1985
authors.initials: KM
authors.lastname: Koo
authors.affiliation: Centre for Personalised Nanomedicine, Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Brisbane, QLD 4072, Australia. j.wee@uq.edu.au yuling.wang@mq.edu.au m.trau@uq.edu.au.
authors.forename: Eugene J H
authors.initials: EJ
authors.lastname: Wee
authors.affiliation: Centre for Personalised Nanomedicine, Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Brisbane, QLD 4072, Australia. j.wee@uq.edu.au yuling.wang@mq.edu.au m.trau@uq.edu.au.
authors.forename: Yuling
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Centre for Personalised Nanomedicine, Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Brisbane, QLD 4072, Australia. j.wee@uq.edu.au yuling.wang@mq.edu.au m.trau@uq.edu.au and School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, QLD 4072, Australia.
authors.forename: Matt
authors.initials: M
authors.lastname: Trau
mesh_terms: D014408:Biomarkers, Tumor / Q000235:genetics; D045744:Cell Line, Tumor; D006801:Humans; D008297:Male; D011471:Prostatic Neoplasms / Q000235:genetics*; D012334:RNA, Neoplasm / Q000235:genetics / Q000652:urine; D012680:Sensitivity and Specificity; D013059:Spectrum Analysis, Raman
chemical_list: D014408:Biomarkers, Tumor; D012334:RNA, Neoplasm