dc.contributor.author | Islam, Md Nazmul | |
dc.contributor.author | Moriam, Sofia | |
dc.contributor.author | Umer, Muhammad | |
dc.contributor.author | Phan, Hoang-Phuong | |
dc.contributor.author | Salomon, Carlos | |
dc.contributor.author | Kline, Richard | |
dc.contributor.author | Nguyen, Nam-Trung | |
dc.contributor.author | Shiddiky, Muhammad JA | |
dc.date.accessioned | 2019-06-19T13:02:57Z | |
dc.date.available | 2019-06-19T13:02:57Z | |
dc.date.issued | 2018 | |
dc.identifier.issn | 0003-2654 | |
dc.identifier.doi | 10.1039/C7AN02109G | |
dc.identifier.uri | http://hdl.handle.net/10072/379864 | |
dc.description.abstract | An 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.peerreviewed | Yes | |
dc.language | English | |
dc.language.iso | eng | |
dc.publisher | Royal Society of Chemistry (RSC) Publications | |
dc.publisher.place | United Kingdom | |
dc.relation.ispartofpagefrom | 3021 | |
dc.relation.ispartofpageto | 3028 | |
dc.relation.ispartofissue | 13 | |
dc.relation.ispartofjournal | Analyst | |
dc.relation.ispartofvolume | 143 | |
dc.subject.fieldofresearch | Analytical chemistry | |
dc.subject.fieldofresearch | Electroanalytical chemistry | |
dc.subject.fieldofresearch | Other chemical sciences | |
dc.subject.fieldofresearchcode | 3401 | |
dc.subject.fieldofresearchcode | 340103 | |
dc.subject.fieldofresearchcode | 3499 | |
dc.title | Naked-eye and electrochemical detection of isothermally amplified HOTAIR long non-coding RNA | |
dc.type | Journal article | |
dc.type.description | C1 - Articles | |
dc.type.code | C - Journal Articles | |
gro.faculty | Griffith Sciences, School of Environment and Science | |
gro.hasfulltext | No Full Text | |
gro.griffith.author | Nguyen, Nam-Trung | |
gro.griffith.author | Shiddiky, Muhammad J. | |
gro.griffith.author | Umer, Muhammad | |