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Article

PCR-Free Detection of Long Non-Coding HOTAIR RNA in Ovarian Cancer Cell Lines and Plasma Samples

1
School of Environment and Science (ESC), Griffith University, Nathan Campus, QLD 4111, Australia
2
Queensland Micro-and Nanotechnology Centre (QMNC), Griffith University, Nathan Campus, QLD 4111, Australia
3
School of Engineering and Built Environment, Griffith University, QLD 4222, Australia
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Section of Gynecologic 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
Department of Clinical Biochemistry and Immunology, Faculty of Pharmacy, University of Concepción, Concepción 4030000, Chile
*
Author to whom correspondence should be addressed.
Cancers 2020, 12(8), 2233; https://doi.org/10.3390/cancers12082233
Received: 24 June 2020 / Revised: 9 July 2020 / Accepted: 4 August 2020 / Published: 10 August 2020
Long non-coding RNA HOX transcript antisense intergenic RNA (HOTAIR) is one of the promising biomarkers that has widely been used in determining the stages of many cancers, including ovarian cancer. In cancer diagnostics, the two key analytical challenges for detecting long non-coding RNA biomarkers are i) the low concentration levels (nM to fM range) in which they are found and ii) the analytical method where broad dynamic range is required (four to six orders of magnitude) due to the large variation in expression levels for different HOTAIR RNAs. To meet these challenges, we report on a biosensing platform for the visual (colorimetric) estimation and subsequent electrochemical quantification of ovarian-cancer-specific HOTAIR using a screen-printed gold electrode (SPE-Au). Our assay utilizes a two-step strategy that involves (i) magnetic isolation and purification of target HOTAIR sequences and (ii) subsequent detection of isolated sequences using a sandwich hybridization coupled with horseradish peroxidase (HRP)-catalyzed reaction of 3,3′,5,5′-tetramethylbenzidine (TMB) in the presence of hydrogen peroxide. The assay achieved a detection limit of 1.0 fM HOTAIR in spiked buffer samples with excellent reproducibility (% RSD ≤ 5%, for n = 3). It was successfully applied to detect HOTAIR in cancer cell lines and a panel of plasma samples derived from patients with ovarian cancer. The analytical performance of the method was validated with standard RT-qPCR. We believe that the proof of concept assay reported here may find potential use in routine clinical settings for the screening of cancer-related lncRNAs. View Full-Text
Keywords: electrochemical detection; HOTAIR RNA; ovarian can cer; naked-eye detection electrochemical detection; HOTAIR RNA; ovarian can cer; naked-eye detection
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MDPI and ACS Style

Soda, N.; Umer, M.; Kashaninejad, N.; Kasetsirikul, S.; Kline, R.; Salomon, C.; Nguyen, N.-T.; Shiddiky, M.J.A. PCR-Free Detection of Long Non-Coding HOTAIR RNA in Ovarian Cancer Cell Lines and Plasma Samples. Cancers 2020, 12, 2233. https://doi.org/10.3390/cancers12082233

AMA Style

Soda N, Umer M, Kashaninejad N, Kasetsirikul S, Kline R, Salomon C, Nguyen N-T, Shiddiky MJA. PCR-Free Detection of Long Non-Coding HOTAIR RNA in Ovarian Cancer Cell Lines and Plasma Samples. Cancers. 2020; 12(8):2233. https://doi.org/10.3390/cancers12082233

Chicago/Turabian Style

Soda, Narshone, Muhammad Umer, Navid Kashaninejad, Surasak Kasetsirikul, Richard Kline, Carlos Salomon, Nam-Trung Nguyen, and Muhammad J.A. Shiddiky 2020. "PCR-Free Detection of Long Non-Coding HOTAIR RNA in Ovarian Cancer Cell Lines and Plasma Samples" Cancers 12, no. 8: 2233. https://doi.org/10.3390/cancers12082233

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