To　overcome　the　dependency　of　strategies　utilizing　cell-free　DNA　(cfDNA)　on　tissue　sampling,　the　emergence　of　sequencing　panels　for　non-invasive　mutation　screening　was　promoted.　However,　cfDNA　sequencing　with　panels　still　suffers　from　either　inaccuracy　or　omission,　and　novel　approaches　for　accurately　screening　tumor　mutations　solely　based　on　plasma　without　gene　panel　restriction　are　urgently　needed.　We　performed　unique　molecular　identifier　(UMI)　target　sequencing　on　plasma　samples　and　peripheral　blood　mononuclear　cells　(PBMCs)　from　85　hepatocellular　carcinoma　(HCC)　patients　receiving　surgical　resection,　which　were　divided　into　an　exploration　dataset　(20　patients)　or　an　evaluation　dataset　(65　patients).　Plasma　mutations　were　identified　in　pre-operative　plasma,　and　the　mutation　variant　frequency　change　(MVFC)　between　post-　and　pre-operative　plasma　was　then　calculated.　In　the　exploration　dataset,　we　observed　that　plasma　mutations　with　MVFC　＜　0.2　were　enriched　for　tumor　mutations　identified　in　tumor　tissues　and　had　frequency　changes　that　correlated　with　tumor　burden;　these　plasma　mutations　were　therefore　defined　as　MVFC-identified　tumor　mutations.　The　presence　of　MVFC-identified　tumor　mutations　after　surgery　was　related　to　shorter　relapse-free　survival　(RFS)　in　both　datasets　and　thus　indicated　minimum　residual　disease　(MRD).　The　combination　of　MVFC-identified　tumor　mutations　and　Alpha　Fetoprotein　(AFP)　could　further　improve　MRD　detection　(P　＜　0.0001).　Identification　of　tumor　mutations　based　on　MVFC　was　also　confirmed　to　be　applicable　with　a　different　gene　panel.　Overall,　we　proposed　a　novel　strategy　for　non-invasive　tumor　mutation　screening　using　solely　plasma　that　could　be　utilized　in　HCC　tumor-burden　monitoring　and　MRD　detection.　©　2023　The　Authors.　Molecular　Oncology　published　by　John　Wiley　&　Sons　Ltd　on　behalf　of　Federation　of　European　Biochemical　Societies.