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Naked-eye and electrochemical detection of isothermally amplified HOTAIR long non-coding RNA
Indexado
WoS WOS:000441407700008
Scopus SCOPUS_ID:85049098605
DOI 10.1039/C7AN02109G
Año 2018
Tipo artículo de investigación

Citas Totales

Autores Afiliación Chile

Instituciones Chile

% Participación
Internacional

Autores
Afiliación Extranjera

Instituciones
Extranjeras


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 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.

Revista



Revista ISSN
Analyst 0003-2654

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Disciplinas de Investigación



WOS
Chemistry, Analytical
Scopus
Sin Disciplinas
SciELO
Sin Disciplinas

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Publicaciones WoS (Ediciones: ISSHP, ISTP, AHCI, SSCI, SCI), Scopus, SciELO Chile.

Colaboración Institucional



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Autores - Afiliación



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

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Financiamiento



Fuente
Fondo Nacional de Desarrollo Científico y Tecnológico
Fondo Nacional de Desarrollo Científico y Tecnológico
National Health and Medical Research Council
Fondo Nacional de Desarrollo Científico, 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

Muestra la fuente de financiamiento declarada en la publicación.

Agradecimientos



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).

Muestra la fuente de financiamiento declarada en la publicación.