Sensitive Detection of Single-Nucleotide Mutation in the BRAF Mutation Site (V600E) of Human Melanoma Using Phosphate–Pyrene-Labeled DNA Probes

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• 作者：Pengfei Li ; Hongyan He ; Zhixuan Wang ; Mengke Feng ; Hongwei Jin ; Yun Wu ; Liangren Zhang ; Lihe Zhang ; Xinjing Tang
• 刊名：Analytical Chemistry
• 出版年：2016
• 出版时间：January 5, 2016
• 年：2016
• 卷：88
• 期：1
• 页码：883-889
• 全文大小：364K
• ISSN：1520-6882

文摘

A series of novel nucleotide phosphoramidites were rationally designed and synthesized and were then site-specifically incorporated in DNA oligonucleotide probes with pyrene-modified phosphate. These oligodeoxynucleotide (ODN) probes almost have no inherent fluorescence emission with pyrene modification at 3′ phosphate of corresponding nucleotides as a result of the photoinduced electron-transfer quenching effect of nucleobases (thymidine ～ cytidine > guanosine ? adenosine). However, strong fluorescence emission was observed only with the perfectly matched duplex for the probes with pyrene modified at 3′ phosphate of thymidine and cytidine. These rationally designed ODN probes successfully worked as “turn on” fluorescence oligonucleotide sensors for single-nucleotide polymorphism (SNP) and were used for detecting a single BRAF mutation site (V600E) of human melanoma.