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
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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.
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