An Interfacial Affinity Interaction-Based Method for Detecting HOTAIR lncRNA in Cancer Plasma Samples
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School of Environment and Science, Nathan Campus, Griffith University, Nathan, QLD 4111, Australia
2
Queensland Micro and Nanotechnology Centre, Nathan Campus, Griffith University, Nathan, QLD 4111, Australia
3
School of Engineering and Built Environment (EBE), Nathan Campus, Griffith University, Nathan, QLD 4111, Australia
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Section of Gynaecologic Oncology, Ochsner Clinic Foundation, New Orleans, LA 70121, USA
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Exosome Biology Laboratory, Centre for Clinical Diagnostics, University of Queensland Centre for Clinical Research, Royal Brisbane and Women’s Hospital, The University of Queensland, Brisbane, QLD 4029, Australia
6
Departamento de Investigación, Postgrado y Educación Continua (DIPEC), Facultad de Ciencias de la Salud, Universidad del Alba, Santiago 8320000, Chile
*
Authors to whom correspondence should be addressed.
Biosensors 2022, 12(5), 287; https://doi.org/10.3390/bios12050287
Received: 30 March 2022
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Revised: 23 April 2022
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Accepted: 26 April 2022
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Published: 28 April 2022
(This article belongs to the Special Issue State-of-the-Art Biosensors in Australia)
Long non-coding RNA Homeobox transcript antisense intergenic RNA (HOTAIR) is recognized as a participant in different processes of normal cell development. Aberrant overexpression of HOTAIR contributes to the initiation, growth, and invasiveness of ovarian cancer. Using the affinity interaction of target HOTAIR lncRNA sequences towards a screen-printed gold electrode (SPE-Au), herein we report on a novel, rapid and simple method to detect HOTAIR sequences. HOTAIR lncRNA sequences were first extracted from ovarian cancer cell lines and patient plasma samples and were magnetically captured and purified by complimentary capture probe-functionalized magnetic beads. Isolated target HOTAIR lncRNAs were directly adsorbed onto unmodified screen-printed gold electrodes (SPE-Au) for direct quantification with [Fe(CN)6]3−/4− redox couple. Our assay achieved a linear dynamic range of 100 nM and 1 pM for detecting pre-clinical model HOTAIR lncRNA samples (%RSD ≤ 5%, for n = 3) and was highly specific, showing clear distinction between HOTAIR lncRNA targets and non-specific miR-891 and miR-486 (100 nM) (%RSD ≤ 5%, for n = 3). The method was tested using ovarian cancer-specific cell lines (SKOV3 and OVCAR3) and mesothelial cell line (MeT-5A)-derived lncRNAs. The analytical performance of our method was validated using RT-qPCR. Finally, the method was tested using clinical samples from ovarian cancer patients and the resulting electrochemical responses show a clear distinction between the ovarian carcinoma and benign samples.
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Keywords:
HOTAIR lncRNA; interfacial biosensing; affinity interaction; RNA biosensor; electrochemical detection; ovarian cancer
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MDPI and ACS Style
Clack, K.; Soda, N.; Kasetsirikul, S.; Kline, R.; Salomon, C.; Shiddiky, M.J.A. An Interfacial Affinity Interaction-Based Method for Detecting HOTAIR lncRNA in Cancer Plasma Samples. Biosensors 2022, 12, 287. https://doi.org/10.3390/bios12050287
AMA Style
Clack K, Soda N, Kasetsirikul S, Kline R, Salomon C, Shiddiky MJA. An Interfacial Affinity Interaction-Based Method for Detecting HOTAIR lncRNA in Cancer Plasma Samples. Biosensors. 2022; 12(5):287. https://doi.org/10.3390/bios12050287
Chicago/Turabian StyleClack, Kimberley, Narshone Soda, Surasak Kasetsirikul, Richard Kline, Carlos Salomon, and Muhammad J.A. Shiddiky. 2022. "An Interfacial Affinity Interaction-Based Method for Detecting HOTAIR lncRNA in Cancer Plasma Samples" Biosensors 12, no. 5: 287. https://doi.org/10.3390/bios12050287
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