GENE MUTATION DETECTION METHOD BASED ON SELECTIVE ELIMINATION OF WILD STRAND BACKGROUND INTERFERENCE

摘要

A gene mutation detection method based on selective elimination of wild strand background interference, comprising: amplifying a target sequence to be detected by means of PCR and then processing the same into single-stranded DNA by means of Lambda exonuclease; designing and synthesizing a thio-DNA strand which is complementary with a target region of a wild type DNA sequence and an RNA closed strand which is complementary with a non-target region, and mixing the same with the single-stranded DNA, and adding DNase I for cutting after temperature rising and annealing; and selectively cutting off a wild type DNA strand target region sequence by the DNase I under the guidance of the thio-DNA strand. A mutation type DNA strand is preserved since the same cannot be cut off by the DNase I because of mispairing in the target region, so that the abundance of the mutation type DNA strand is remarkably improved and the difficulty in detecting low-abundance gene mutation is greatly reduced. The method disclosed by the present invention does not need complicated and expensive instruments, is easy to operate and low in cost and may provide a result within one day. In-time and reliable gene mutation information may be provided for clinical early screening of tumors and monitoring of recurrence after operation.