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Citation: WEI Jia, GAO Jia-Xue, WANG Yao-Qi, WANG Zhen-Xin, MENG Xian-Ying. Specific and Sensitive Detection of BRAFV600E Mutations in Papillary Thyroid Carcinoma by Oligonucleotide Microarray-based Fluorescence Assay[J]. Chinese Journal of Analytical Chemistry, ;2019, 47(6): 855-861. doi: 10.19756/j.issn.0253-3820.191149 shu

Specific and Sensitive Detection of BRAFV600E Mutations in Papillary Thyroid Carcinoma by Oligonucleotide Microarray-based Fluorescence Assay

1.
Department of Thyroid Surgery, the First Hospital of Jilin University, Changchun 130021, China;
2.
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China

Received Date: 20 March 2019

Fund Project: This work was supported by the Changchun Science and Technology Project (No. 18YJ009) and the National Natural Science Foundation of China (No. 21775145).

A DNA microarray-based fluorescence assay was developed for profiling BRAFV600 mutations through formation of three single-stranded DNA (ssDNA) sequences hybridization systems on microarray. In this assay, the target ssDNAs were recognized by correspondent capturing oligonucleotide probes on microarray, followed by hybridization with a Cy5 modified ssDNA for fluorescence detection. Under the optimal experimental conditions, the DNA microarray-based fluorescence assay achieved a limit of detection (S/N) at sub-nanomolar level for the target ssDNA with BRAFV600E mutation, and could easily differentiate as low as 0.1% BRAFV600E mutation sequence in the mixture of target ssDNAs. The profile results of BRAFV600E mutation level in 15 clinical thyroid tissue samples demonstrated that the DNA microarray-based fluorescence assay had good practicability.
- Papillary thyroid carcinoma,
- BRAFV600E mutations,
- DNA microarray
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