title: High-speed biosensing strategy for non-invasive profiling of multiple cancer fusion genes in urine.
abstract: Aberrant chromosal rearrangements, such as the multiple variants of TMPRSS2:ERG fusion gene mutations in prostate cancer (PCa), are promising diagnostic and prognostic biomarkers due to their specific expression in cancerous tissue only. Additionally, TMPRSS2:ERG variants are detectable in urine to provide non-invasive PCa diagnostic sampling as an attractive surrogate for needle biopsies. Therefore, rapid and simplistic assays for identifying multiple urinary TMPRSS2:ERG variants are potentially useful to aid in early cancer detection, immediate patient risk stratification, and prompt personalized treatment. However, current strategies for simultaneous detection of multiple gene fusions are limited by tedious and prolonged experimental protocols, thus limiting their use as rapid clinical screening tools. Herein, we report a simple and rapid gene fusion strategy which expliots the specificity of DNA ligase and the speed of isothermal amplification to simultaneously detect multiple fusion gene RNAs within a short sample-to-answer timeframe of 60min. The method has a low detection limit of 2 amol (1000 copies), and was successfully applied for non-invasive fusion gene profiling in patient urine samples with subsequent validation by a PCR-based gold standard approach.
authors.affiliation: Centre for Personalized Nanomedicine, Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, QLD 4072, Australia.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Koo
authors.affiliation: Centre for Personalized Nanomedicine, Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, QLD 4072, Australia. Electronic address: j.wee@uq.edu.au.
authors.forename: Eugene J H
authors.initials: EJH
authors.lastname: Wee
authors.affiliation: Centre for Personalized Nanomedicine, Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, QLD 4072, Australia; School of Chemistry and Molecular Biosciences, The University of Queensland, QLD 4072, Australia. Electronic address: m.trau@uq.edu.au.
authors.forename: Matt
authors.initials: M
authors.lastname: Trau
mesh_terms: D014408:Biomarkers, Tumor / Q000235:genetics / Q000652:urine; D015374:Biosensing Techniques / Q000191:economics / Q000379:methods; D006801:Humans; D008297:Male; D021141:Nucleic Acid Amplification Techniques / Q000191:economics / Q000379:methods*; D050596:Oncogene Fusion; D015514:Oncogene Proteins, Fusion / Q000235:genetics* / Q000652:urine*; D011467:Prostate / Q000473:pathology; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000652:urine*; D013050:Spectrometry, Fluorescence / Q000379:methods; D013997:Time Factors; D059467:Transcriptome
chemical_list: D014408:Biomarkers, Tumor; D015514:Oncogene Proteins, Fusion; C515553:TMPRSS2-ERG fusion protein, human
keywords: Gene fusion biomarkers; Non-invasive detection; Prostate cancer cell lines; Recombinase polymerase amplification; Urine samples