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A fluorescence method for homogeneous detection of influenza A DNA sequence based on guanine‐quadruplex‐N‐methylmesoporphyrin IX complex and assistance‐DNA inhibition

机译:基于鸟嘌呤 - Quadreple-N-甲基甲基卟啉IX复合物和辅助-DNA抑制的流感均匀检测荧光方法

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摘要

Abstract In his study, we report a fluorescence method for homogeneous detection of influenza A (H1N1) DNA sequence based on G‐quadruplex‐NMM complex and assistance‐DNA (A‐DNA) inhibition. The quadruplex‐based functional DNA (QBF‐DNA), composed of a complementary probe to the target H1N1 DNA sequence and G‐rich fragment, was designed as the signal DNA. The A‐DNA consisted of two parts, one part was complementary to target H1N1 DNA and the other part was complementary to the signal DNA. In the absence of target H1N1 DNA, the G‐rich fragment of QBF‐DNA can form G‐quadruplex‐NMM complex, which outputted a fluorescent signal. With the presence of target H1N1 DNA, QBF‐DNA, and A‐DNA can simultaneously hybridize with target H1N1 DNA to form double‐helix structure. In this case, the A‐DNA partially hybridized with the QBF‐DNA, which inhibited the formation of G‐quadruplex‐NMM complex, leading to the decrease of fluorescent signal. Under the optimum conditions, the fluorescence intensity was inversely proportional to the concentration of target H1N1 DNA over the range from 25 to 700 pmol/L with a detection limit of 8 pmol/L. In addition, the method is target specific and practicability, and would become a new diagnostic assay for H1N1 DNA sequence and other infectious diseases.
机译:摘要在他的研究中,我们报告了一种基于G-Quadreple-NMM复合物和辅助-DNA(A-DNA)抑制的流感A(H1N1)DNA序列的均匀检测荧光方法。设计由互补探针与富含靶H1N1 DNA序列和富含G的片段组成的Quadreplex的官能DNA(QBF-DNA)作为信号DNA。 A-DNA由两部分组成,一个部分与靶H1N1 DNA互补,另一部分与信号DNA互补。在没有靶H1N1 DNA的情况下,富QBF-DNA的富型片段可以形成G-Quadreple-NMM复合物,其输出荧光信号。随着靶H1N1 DNA,QBF-DNA和A-DNA的存在,可以与靶H1N1 DNA同时杂交以形成双螺旋结构。在这种情况下,与QBF-DNA部分杂交的A-DNA抑制了G-Quadreple-NMM复合物的形成,导致荧光信号的降低。在最佳条件下,荧光强度与靶H1N1 DNA的浓度与25至700pmol / L的浓度成反比,具有8pmol / L的检测限。此外,该方法是目标特异性和实用性,并且将成为H1N1 DNA序列和其他传染病的新诊断测定。

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