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| DOI | 10.1039/C7AN02109G | ||||
| Año | 2018 | ||||
| Tipo | artículo de investigación |
Citas Totales
Autores Afiliación Chile
Instituciones Chile
% Participación
Internacional
Autores
Afiliación Extranjera
Instituciones
Extranjeras
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 noncoding HOX transcript antisense intergenic RNA (HOTAIR) detection. Isothermal reverse transcription-recombnase 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.
| Ord. | Autor | Género | Institución - País |
|---|---|---|---|
| 1 | Islam, Md Nazmul | - |
Griffith Univ - Australia
Griffith University - Australia |
| 2 | Moriam, Sofia | Mujer |
Griffith Univ - Australia
Griffith University - Australia |
| 3 | Umer, Muhammad | Hombre |
Griffith Univ - Australia
Griffith University - Australia |
| 4 | Phan, Hoang-Phuong | Hombre |
Griffith Univ - Australia
Griffith University - Australia |
| 5 | SALOMON-GALLO, CARLOS FRANCISCO | Hombre |
UNIV QUEENSLAND - Australia
Universidad de Concepción - Chile Ochsner Clin Fdn - Estados Unidos University of Queensland, Centre for Clinical Research - Australia Ochsner Health System - Estados Unidos UQ Centre for Clinical Research - Australia Ochsner Health - Estados Unidos |
| 6 | Kline, Richard | Hombre |
Ochsner Clin Fdn - Estados Unidos
Ochsner Health System - Estados Unidos Ochsner Health - Estados Unidos |
| 7 | Nguyen, Nam-Trung | Hombre |
Griffith Univ - Australia
Griffith University - Australia |
| 8 | Shiddiky, Muhammad J. A. | Hombre |
Griffith Univ - Australia
Griffith University - Australia |
| Fuente |
|---|
| Fondo Nacional de Desarrollo Científico y Tecnológico |
| Fondo Nacional de Desarrollo CientÃfico y Tecnológico |
| National Health and Medical Research Council |
| Fondo Nacional de Desarrollo CientÃfico, Tecnológico y de Innovación Tecnológica |
| Lions Medical Research Foundation |
| Ovarian Cancer Research Foundation |
| Griffith University |
| NHMRC CDF |
| Lions Medical Research Foundation (LMRF) |
| Ovarian Cancer Research Foundation (OCRF) |
| GUPRS |
| GUIPRS |
| Canadian Dermatology Foundation |
| Agradecimiento |
|---|
| This work was supported by the NHMRC CDF (APP1088966 to M. J. A. S.) and higher degree research scholarships (GUIPRS and GUPRS) to M. N. I. from the Griffith University. Dr Carlos Salomon is supported by The Lions Medical Research Foundation (LMRF), Ovarian Cancer Research Foundation (OCRF) and Fondo Nacional de Desarrollo Cientifico y Tecnologico (FONDECYT 1170809). |
| This work was supported by the NHMRC CDF (APP1088966 to M. J. A. S.) and higher degree research scholarships (GUIPRS and GUPRS) to M. N. I. from the Griffith University. Dr Carlos Salomon is supported by The Lions Medical Research Foundation (LMRF), Ovarian Cancer Research Foundation (OCRF) and Fondo Nacional de Desarrollo Científico y Tecnológico (FONDECYT 1170809). |