High-
throughput detection for the interesting base or the
mutation site in the
nucleic acid sample can be achieved by the linear
test probe pairs P1 and P2. The
test probe pairs P1 and P2 respectively comprise either of the flanking complementary sequences which are adjacent to the interesting base or the
mutation site in the
nucleic acid sample. When the
test probe pairs P1, P2 are annealed and hybridized to the
nucleic acid sample, a gap will be generated at the interesting base or the
mutation site position between the probe pairs and the sample. Divide the annealed
hybrid sample into four equal reaction systems to which add dATP, dTTP, dCTP, dGTP, respectively. The test probe pairs P1 and P2 will be ligated into one
single probe when adding the complementary
nucleotide system under the
DNA polymerase or ligase. After purified and amplified, the generated single probes are hybridized to the corresponding area in a common
oligonucleotide microarray. The generated
single probe will give a
signal in the
hybrid area, and therefore detect and analyze the
hybrid signal to determine the base type or the mutation
genotype at the detection position. The invention can be applied to the re-sequencing the target
nucleic acid sequence, the detection and analysis for the mutation,
insertion, or deletion sites of a known
nucleic acid sequence, and the
genotyping of the
pathogenic microorganism.