The invention relates to a method and a kit for enriching 4000 human pathogenic target genes. The method comprises the following steps: breaking
DNA to obtain
DNA fragments, carrying out terminal repairing, carrying out A addition, carrying out
linker addition, and carrying out Pre-Capture LM-PCR; mixing Pre-Capture LM-PCR products of 4000 human pathogenic target genes to obtain a mixed
library sample, hybridizing a specific probe
library of the 4000 human pathogenic target genes with the mixed
library sample, capturing
DNA by adopting magnetic beads, and carrying out eluting; and carrying out Post-Capture LM-PCR. The kit contains
a DNA terminal repairing
reagent, an A addition
reagent, a
linker addition
reagent, an LM-PCR reagent and a capturing reagent. According to the method and the kit, the 4000 human
pathogenic genes are taken as target spots and are combined together, the
gene sequences are enriched once through a
biotin probe capturing technology and are subjected to high-
throughput sequencing, therefore, multiple variation types of the genes can be simultaneously examined, and the detection sensitivity is high; the 4000
pathogenic genes can be taken as a principal diagnosis and detection means clinically, the cost is reduced, the
missed diagnosis is reduced, the detection positive rate is improved, and therefore, the clinical popularization is easy.