| 摘要 |
<p>The present invention provides an improved method of detecting the presence of a nucleotide sequence within a double-stranded DNA in a sample comprising the following steps: a) coating a solid support with a first layer of biotinylated serum albumin, and a second layer of streptaviding having sufficient density to perform efficient microarray analysis; b) digesting the double-stranded DNA with an exonuclease to convert double-stranded to single stranded DNA, derived from a mixture of target cells and other cells, to a single-stranded DNA; c) capturing a first nucleic acid probe adapted by biotin to said coated solid support defined in step a; d) hybridizing (i) the single-stranded DNA with the first nucleic acid probe, and (ii) a second nucleic acid probe labeled with a detectable moiety which can hybridize with the single-stranded DNA adjacent the hybridized first nucleic probe; e) ligating the hybridized first and second nucleic acid probes in case of perfect match only; f) denaturing the ligated first and second nucleic acid probes from the hybridized single-stranded DNA; g) removing non-covalently bound labeled probes and single stranded DNA; and h) detecting captured detectable moiety indicating the presence of the nucleotide sequence within the double-stranded DNA in the sample; wherein steps c.-h. are performed by microarray technique. Also provided is a device and a kit suitable for carrying out said detection method.</p> |
