High-throughput quantitative detection of triple-negative breast cancer-associated expressed miRNAs by rolling circle amplification on fluorescence-encoded microspheres |
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| Institution: | 1. Key Laboratory of Tropical Translational Medicine of Ministry of Education, NHC Key Laboratory of Control of Tropical Diseases, School of Tropical Medicine, Micronano Technology Research Institute, School of Basic Medicine and Life Sciences, Hainan Medical University, Haikou 571199, China;2. Beadstar Biotechnology Hainan Province Co., Ltd., Haikou 570110, China;3. State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China;4. Economical Forest Cultivation and Utilization of 2011 Collaborative Innovation Center in Hunan Province, Hunan Key Laboratory of Biomedical Nanomaterials and Devices, Hunan University of Technology, Zhuzhou 412007, China;1. Liaoning & Shenyang Key Laboratory of Functional Dye and Pigment, Shenyang University of Chemical Technology, Shenyang 110142, China;2. Department of Cell Biology, China Medical University, Shenyang 110122, China;3. State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 110624, China;4. Department of Chemistry, Graduate School of Science, Hiroshima University, Higashi-Hiroshima 7398526, Japan;1. College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China;2. State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry & Chemical Engineering and Center of Materials Analysis, Nanjing University, Nanjing 210023, China;3. Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing 210023, China;1. Changsha Medical University, Academician Workstation, Changsha 410219, China;2. Xiangya School of Pharmaceutical Sciences, Central South University, Changsha 410013, China;3. State Key Laboratory of Toxicology and Medical Countermeasures, Institute of Pharmacology and Toxicology, Beijing 100850, China;4. Key Laboratory of Biological Nanotechnology of National Health Commission, Changsha 410000, China |
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| Abstract: | Compared with other types of breast cancer, triple-negative breast cancer (TNBC) has the characteristics of a high degree of malignancy and poor prognosis. Early diagnosis of TNBC through biological markers and timely development of effective treatment methods can reduce its mortality. Many Research experiments have confirmed that some specific miRNA expression profiles in TNBC can used as markers for early diagnosis. However, detecting the expression profiles of multiple groups of miRNAs according to traditional detection methods is complicated and consumes many samples. To address this issue, we developed a method for high-throughput, high-sensitivity quantitative detection of multiple sets of miRNAs (including miR-16, miR-21, miR-92, miR-199, and miR-342) specifically expressed in TNBC by rolling circle amplification (RCA) on fluorescence-encoded microspheres. Through the optimization of reaction system conditions, the developed method showed an extensive linear dynamic range and high sensitivity for all five miRNAs with the lowest limit of detection of 2 fmol/L. Meanwhile, this high-throughput detection method also appeared reasonable specificity. Only in the presence of a specific target miRNA, the fluorescence signal on the correspondingly encoded microspheres is significantly increased, while the fluorescence signal on other non-correspondingly encoded microspheres is almost negligible. Furthermore, this process exhibited good recovery and reproducibility in serum. The advantages of this method allow us to more conveniently obtain the expression profiles of multiple groups of TNBC-associated miRNAs, which is beneficial for the early detection of TNBC. |
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