The invention discloses a next generation sequencing-based detection panel and detection kit for pan-cancer targeting, chemotherapy and immune drugs and application thereof. The detection panel includes gene mutations related to pan-cancer type, treatment and prognosis, tumor mutation load calculation related exon regions and microsatellite instability sites. According the technical solution of the present invention, the detection panel includes the gene mutations related to pan-cancer type, treatment and prognosis, the tumor mutation load calculation related exon regions and the microsatellite instability sites, and the gene information included is comprehensive; a variety of tumor mutations can be jointly detected; and the detection panel and detection kit can be used for the concomitantdiagnosis of targeted drugs, chemotherapeutics or immune drugs to obtain accurate results.