Provided are a method and a combination for one-tube simultaneous detection of tumor-specific
gene mutation and
methylation, and the use thereof. The method comprises searching a
database to select
mutation and
methylation sites of a
cancer driver
gene and a
cancer suppressor gene, synthesizing capture probes correspondingly paired with a target interval, performing
hybrid capture on an amplification product of a sample treated with a restriction
endonuclease by using a probe set comprising tumor-specific
gene mutation and
methylation capture probes, constructing a high-
throughput sequencing
library containing a target region, and performing high-
throughput sequencing to obtain the sequence of a target fragment, so as to obtain the results of methylation, point mutations and
indel variation of a
cancer-associated gene. The provided method enables one-tube simultaneous detection of
mutation and methylation states of a cancer-associated gene in one
assay, requires a low content of a gene to be detected, can be used for methylation and
mutation detection of a tumor-specific
DNA, and has low detection costs and high efficiency.