Detection of FGFR2:FAM76A Fusion Gene in Circulating Tumor RNA Based on Catalytic Signal Amplification of Graphene Oxide-loaded Magnetic Nanoparticles,Electroanalysis

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Detection of FGFR2:FAM76A Fusion Gene in Circulating Tumor RNA Based on Catalytic Signal Amplification of Graphene Oxide-loaded Magnetic Nanoparticles
Electroanalysis ( IF 2.7 ) Pub Date : 2018-07-04 , DOI: 10.1002/elan.201800282
Lena Gorgannezhad _{1,

2} , Muhammad Umer ₂ , Mostafa Kamal Masud _{2,

3} , Md. Shahriar A. Hossain ₃ , S. Tanaka ₄ , Y. Yamauchi _{3,

4,

5} , Carlos Salomon _{6,

7,

8} , Richard Kline ₈ , Nam-Trung Nguyen ₂ , Muhammad J. A. Shiddiky _{1,

2}

Affiliation

School of Environment and Science; Griffith University, Nathan Campus; QLD 4111 Australia
Queensland Micro- and Nanotechnology Centre; Griffith University, Nathan Campus; QLD 4111 Australia
School of Chemical Engineering, School of Mechanical and Mining Engineering, and Australian Institute for Bioengineering and Nanotechnology (AIBN); The University of Queensland; Brisbane QLD 4072 Australia
International Center for Materials Nanoarchitectonics (WPI-MANA); National Institute for Materials Science (NIMS); 1-1 Namiki, Tsukuba Ibaraki 305-0044 Japan
Department of Plant and Environmental New Resources; Kyung Hee University; 1732 Deogyeong-daero Giheung-gu, Yongin-si, Gyeonggi-do 446-701 South Korea
Exosome Biology Laboratory, Centre for Clinical Diagnostics, University of Queensland Centre for Clinical Research, Royal Brisbane and Womens Hospital; The University of Queensland; Brisbane, QLD Australia
Department of Clinical Biochemistry and Immunology, Faculty of Pharmacy; University of Concepción; Concepción Chile
Maternal-Fetal Medicine, Department of Obstetrics and Gynecology; Ochsner Clinic Foundation; New Orleans USA

Circulating tumor nucleic acids (ctNAs) are promising biomarkers for minimally invasive cancer assessment. The FGFR2 : FAM76A fusion gene is one of the highly promising ovarian cancer biomarkers detectable in ctNAs. Herein, we introduce a new amplification-free electrochemical assay for the detection of FGFR2 : FAM76A fusion gene in ctNAs extracted from ovarian cancer patients. The assay relies on the electrocatalytic activity of a new class of superparamagnetic graphene-loaded iron oxide nanoparticles (GO-NPFeO). After isolation and purification, the target RNA was directly adsorbed onto the GO-NPFeO surface through graphene-RNA affinity interaction. The electrocatalytic signal was achieved by the reduction of surface-attached ruthenium hexaammine(III) chloride which was further amplified by using the ferricyanide redox system. Our assay depicted an excellent detection sensitivity down to 1.0 fM, high specificity and excellent reproducibility (% RSD=<5 %, for n=3). The analytical performance of our method was validated with standard qRT-PCR analysis. We believe that this newly developed assay would be practically applicable in clinical research.

中文翻译：

基于氧化石墨烯磁性纳米粒子催化信号放大检测循环肿瘤RNA中FGFR2:FAM76A融合基因

循环肿瘤核酸 (ctNA) 是用于微创癌症评估的有前途的生物标志物。FGFR2 : FAM76A 融合基因是可在 ctNA 中检测到的非常有希望的卵巢癌生物标志物之一。在此，我们介绍了一种新的无扩增电化学检测方法，用于检测从卵巢癌患者中提取的 ctNA 中的 FGFR2：FAM76A 融合基因。该测定依赖于一类新型超顺磁性石墨烯负载氧化铁纳米粒子 (GO-NPFeO) 的电催化活性。分离纯化后，目标RNA通过石墨烯-RNA亲和相互作用直接吸附到GO-NPFeO表面。电催化信号是通过还原表面附着的六氨合氯化钌 (III) 来实现的，该信号通过使用铁氰化物氧化还原系统进一步放大。我们的分析显示出低至 1.0 fM 的出色检测灵敏度、高特异性和出色的重现性（% RSD=<5%，对于 n=3）。我们方法的分析性能通过标准 qRT-PCR 分析进行了验证。我们相信这种新开发的检测方法将实际适用于临床研究。

更新日期：2018-07-04

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