The invention provides a plasma cfDNA (cell-free deoxyribonucleic acid) bi-molecular marker. The bi-molecular marker is oligonucleotide, the sequence is represented as follows: a) 5'P-GACGTC-GATCGGAAGAGCTCGTATGCCGTCTTCTGCTTG, b) 5' ACACTCTTTCCCTACACGACGCTCTTCCGATCT-NNNNNNNNNNNNNNNNNNNN-GACGTCT, the sequence of oligonucleotide is from the 5' terminal to the 3' terminal from left to right, N represents a random basic group, and P represents a phosphate group; the oligonucleotide sequence is synthesized artificially. The plasma cfDNA bi-molecular marker is used for distinguishing every plasma cfDNA molecule by adding a unique molecular marker to every plasma cfDNA molecule and can be applied to plasma cfDNA detection. The plasma cfDNA bi-molecular marker overcomes defects of cfDNA detection with a conventional NGS (next generation sequencing) technology, and specifically has following technical effects: 1), the cfDNA valid data volume in a database construction process is increased; 2), noise signals such as deviation and detection errors produced in intermediate steps of gene detection can be reduced; 3), target variation can be effectively detected, meanwhile, low false positive rate is realized, and accuracy and stability of gene detection are improved.