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

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 wit...

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Bibliographic Details
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
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Summary: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.
ISSN:0003-2654
1364-5528
DOI:10.1039/d0an02074e