A novel FRET biosensor based on four-way branch migration HCR for Vibrio parahaemolyticus detection
A novel FRET biosensor based on four-way branch migration HCR for Vibrio parahaemolyticus detection
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DOI：
10.1016/j.snb.2019.05.054
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发表时间：
2019-10
期刊：
影响因子：
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通讯作者：
Dong-xia Ren;Chengjun Sun;Zhijun Huang;Zewei Luo;Chen Zhou;Yongxin Li
中科院分区：
文献类型：
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作者：
Dong-xia Ren;Chengjun Sun;Zhijun Huang;Zewei Luo;Chen Zhou;Yongxin Li
提出了一种基于四路分支迁移杂交链式反应（HCR）的多功能、强大的荧光共振能量转移（FRET）生物传感器，用于副溶血弧菌（V. parahaemolyticus）的检测。在该系统中，设计了两个部分互补的H1和H2发夹结构以及一条辅助DNA链（R）。 H1 用羧基荧光素 (FAM) 标记作为荧光团供体，H2 用四甲基罗丹明 (TAMRA) 标记作为荧光团受体。当目标DNA出现在系统中时，它首先与辅助DNA结合形成短的双链DNA（dsDNA），随后触发四向分支迁移HCR，产生长切口的dsDNA多联体，并使大量的FAM和TAMRA靠近，从而获得显着的FRET信号。借助精心设计的四路分支迁移HCR电路，该FRET生物传感器表现出卓越的检测能力，可以检测0.067nM的目标DNA和低至10 CFU mL−1ofV。副溶血性杆菌。据我们所知，这是首次将用于信号放大的四向分支迁移HCR策略应用于FRET生物传感器，这可能在食品安全和临床诊断方面显示出巨大的潜力。
A versatile and powerful fluorescence resonance energy transfer (FRET) biosensor based on four-way branch migration hybridization chain reaction (HCR) was proposed for the detection ofVibrio parahaemolyticus(V. parahaemolyticus). In the system, two partly complementary hairpin structures of H1 and H2, and an assistant DNA strand (R) were designed. H1 was labeled with carboxyfluorescein (FAM) as fluorophore donor, and H2 was labeled with tetramethylrhodamine (TAMRA) as fluorophore acceptor. When target DNA was presented in the system, it would firstly bind with the assistant DNA to form a short double-strand DNA (dsDNA), and subsequently trigger the four-way branch migration HCR, which would produce long nicked dsDNA concatamer and bring a good deal of FAM and TAMRA in close proximity, so that dramatic FRET signals could be achieved. With the assistance of well-designed four-way branch migration HCR circuits, this FRET biosensor exhibited a superior detection capability, which could detect 0.067 nM target DNA and as low as 10 CFU mL−1ofV. parahaemolyticus. To our knowledge, it is the first time that a four-way branch migration HCR strategy for signal amplification has been adapted to the FRET biosensor, which might show great potential in food safety and clinical diagnosis.