A versatile magnetic bead-based flow cytometric assay for the detection of thyroid cancer related hsa-miR-221-3p in blood and tissues

• Published in: Analyst (London) 2021-02, Vol.146 (3), p.842-847
• Main Authors: Gao, Jiaxue, Wei, Jia, Wang, Yaoqi, Li, Zhenshengnan, Ma, Lina, Meng, Xianying, Wang, Zhenxin
• Format: Article
• Language: English
• Subjects: Assaying; Biomolecules; Cancer; Deoxyribonucleic acid; Diagnosis; DNA; Flow Cytometry; Fluorescence; Humans; Magnetic Phenomena; MicroRNAs - genetics; Ribonucleic acid; RNA; Room temperature; Thyroid cancer; Thyroid Cancer, Papillary - genetics; Thyroid Neoplasms - diagnosis; Thyroid Neoplasms - genetics
• ISSN: 0003-2654; 1364-5528
• DOI: 10.1039/d0an02074e

In vitro detection of low abundance biomolecules including microRNAs (miRNAs) is essential to biological research and early clinical diagnosis. In this work, a versatile magnetic bead (MB)-based flow cytometric assay was developed for the detection of hsa-miR-221-3p, which is strongly associated with papillary thyroid carcinoma (PTC). In the presence of hsa-miR-221-3p, the complementary DNA probe attached to the surface of MBs is hybridized with the target to form DNA/RNA heteroduplexes. After the recognition of the DNA/RNA heteroduplexes by PicoGreen, the fluorescence signals of each MB were readily detected using a flow cytometer. This assay can selectively detect hsa-miR-221-3p with a detection limit of 2.1 pM. The practicality of the assay is demonstrated by the discrimination of thyroid cancer tissues from normal tissues, and a satisfactory result is obtained. Moreover, this assay can be rapidly carried out in one step at room temperature, providing a generic method for the sensitive detection of miRNAs in molecular diagnosis. An MB-FCM assay for the sensitive detection of hsa-miR-221-3p in native biological samples in one step at room temperature.