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Abstract

This invention relates to a pair of probes for the detection of a target nucleic acid, said pair of probes comprising (1) probe 1 having base sequence B1 complementary to A1 between two specific sequential base sequences A1 and A2 of the nucleic acid, wherein (a) the 5'-terminus of base sequence B1 is phosphorylated and (b) a first solid phase immobilizing part is bound to the 3'-terminus of base sequence B1, and (2) probe 2 having base sequence B2 complementary to A2 between base sequences A1 and A2, wherein (a) the 5'-terminus of base sequence B2 is bound to one end of a cleavage part and (b) a second solid phase immobilizing part is bound to the other part of the cleavage part. When the target nucleic acid is hybridized above the solid phase of the invention where said probes are immobilized on its surface, the probes on the solid phase occupy spatial positions beneficial to the formation of a hybrid and thus forms the hybrid efficiently. Therefore, probe 1 and probe 2 are efficiently ligated by ligase reaction. Furthermore, after the target nucleic acid is removed from the hybrid, only probe 2 ligated as described above will be able to exist on the solid phase through cleavage reaction of the cleavage part. Therefore, after the free probe 2 has been removed by washing, it will become possible to detect the presence of probe 2 on the solid phase ligated as described above, with high sensitivity and high recognition. This will enable detection of the presence of a target nucleic acid with higher sensitivity and higher recognition as compared to methods in the prior art.

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First Claim