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dc.contributor.authorIslam, Md Nazmul
dc.contributor.authorMoriam, Sofia
dc.contributor.authorUmer, Muhammad
dc.contributor.authorPhan, Hoang-Phuong
dc.contributor.authorSalomon, Carlos
dc.contributor.authorKline, Richard
dc.contributor.authorNguyen, Nam-Trung
dc.contributor.authorShiddiky, Muhammad JA
dc.date.accessioned2019-06-19T13:02:57Z
dc.date.available2019-06-19T13:02:57Z
dc.date.issued2018
dc.identifier.issn0003-2654
dc.identifier.doi10.1039/C7AN02109G
dc.identifier.urihttp://hdl.handle.net/10072/379864
dc.description.abstractAn inexpensive, simple and rapid sensor platform capable of detecting cancer-related long non-coding RNA (lncRNA) with high accuracy is of great interest in the field of molecular diagnostics. Herein, we report on the development of a new colorimetric and electrochemical assay platform for long non-coding HOX transcript antisense intergenic RNA (HOTAIR) detection. Isothermal reverse transcription-recombinase polymerase amplification (RT-RPA) was performed to amplify HOTAIR sequences from a RNA pool extracted from a designated number of ovarian cancer cells and a small cohort of plasma samples derived from patients with ovarian cancer. During RT-RPA, biotinylated dUTPs were randomly incorporated in the amplified product. Subsequently, HOTAIR amplicons were magnetically purified and isolated followed by a horseradish peroxidase (HRP)-catalyzed colorimetric reaction in the presence of the 3,3′,5,5′-tetramethylbenzidine (TMB)/H2O2 system. We finally introduced three potential readout methods for HOTAIR detection – (i) naked-eye visualisation of the color change for a quick screening of the target, (ii) quantitative absorbance measurement by UV-vis, and (iii) amperometric quantification using the electrochemical properties of TMB. The assay has shown excellent reproducibility (% RSD = <5%, for n = 3) and sensitivity (10 cells/ per mL) while detecting HOTAIR in cancer cell lines and patient samples. The expression of HOTAIR in clinical samples was also verified with a standard RT-qPCR method. We believe that our proof of concept assay may find potential relevance for the routine clinical screening of cancer-associated lncRNAs.
dc.description.peerreviewedYes
dc.languageEnglish
dc.language.isoeng
dc.publisherRoyal Society of Chemistry (RSC) Publications
dc.publisher.placeUnited Kingdom
dc.relation.ispartofpagefrom3021
dc.relation.ispartofpageto3028
dc.relation.ispartofissue13
dc.relation.ispartofjournalAnalyst
dc.relation.ispartofvolume143
dc.subject.fieldofresearchAnalytical chemistry
dc.subject.fieldofresearchElectroanalytical chemistry
dc.subject.fieldofresearchOther chemical sciences
dc.subject.fieldofresearchcode3401
dc.subject.fieldofresearchcode340103
dc.subject.fieldofresearchcode3499
dc.titleNaked-eye and electrochemical detection of isothermally amplified HOTAIR long non-coding RNA
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
gro.facultyGriffith Sciences, School of Environment and Science
gro.hasfulltextNo Full Text
gro.griffith.authorNguyen, Nam-Trung
gro.griffith.authorShiddiky, Muhammad J.
gro.griffith.authorUmer, Muhammad


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