Patent ID: 6821734
Filing Date: 2004-11-23
Classification: C12Q

Abstract:
A method for examining nucleotide sequences which comprises:(1) adding a group of primers consisting of multiple primer species to a solution containing a sample of nucleic acid subjected to examination, and performing simultaneous synthesis of complementary strands at each of the multiple regions of the nucleic acid containing target nucleotide sequences to be examined; (2) designing DNA probes with specific sequences so that elongation of complementary strands is affected by the presence or absence of mutations in said target nucleotide sequences wherein the same number of such DNA probes and said target sequences is used for elongation of complementary strands; (3) immobilizing said DNA probes in subcells of a reaction vessel that are compartmentalized for each said DNA probe; (4) adding the solution obtained after step (1) to the reaction vessel, wherein said DNA probes are allowed to hybridize with said target sequences or sequences complementary to said target sequences under condition that the solution can freely move among said subcells, and then the excess amount of the solution is removed so that the remaining solution can no longer freely move among said subcells; (5) performing elongation reaction of complementary strands using said target sequences or the sequences complementary to said target sequences as a template and the following reaction, in which pyrophosphate produced during said elongation reaction is converted to ATP and reacted with chemiluminescent substrates to develop luminescence, in the subcells of the reaction vessel, wherein elongation of complementary strand occurs for the nucleic acid whose sequences are complementary to said probes; and (6) determining the presence or absence of mutations in said target nucleotide sequences based upon said luminescence.