This invention discloses a hexavanadate-bridged double-ended click
nucleic acid probe construct, its preparation method, and its application, relating to the field of bioanalytical technology. The hexavanadate-bridged double-ended click
nucleic acid probe construct comprises a hexavanadate derivative V6-(N3)2 bridging
scaffold with azido groups at both ends, and two identical oligonucleotides MB1 or MB2; MB1 is terminally
alkyne and labeled with a FAM fluorescent group, and MB2 is terminally
alkyne and labeled with a Cy5 fluorescent group; V6-(N3)2 and MB1 or MB2 undergo a Cu(I)-catalyzed azido-alkynyl
cycloaddition reaction to form a 1,2,3-
triazole linker bond, connecting to form an MB1-
triazole-V6-
triazole-MB1 or MB2-triazole-V6-triazole-MB2 structure. This invention also discloses a method for preparing this construct, wherein the finished product is obtained through a click reaction in an
aqueous buffer system,
copper removal and desalting, and separation and purification. And its application in
nucleic acid sequence-
specific detection, with its
fluorescence quantitative readout mechanism, improves the robustness of sequence identification, mismatch detection and quantitative detection, and is suitable for
pathogen identification,
gene mutation screening,
molecular diagnostics and
drug companion diagnostics.