The invention discloses a kit and method for detecting chromosome heterozygosity loss on the basis of amplicon sequencing, belongs to the field of molecular biology nucleic acid detection, and relatesto the kit and method for detecting chromosome heterozygosity loss by means of amplicon sequencing. The kit contains a mixture of 14 groups of primer pairs used for detecting a 1p36.32 region of a human chromosome 1, 6 groups of primer pairs used for detecting a 1q25.2 region of the human chromosome 1, 14 groups of primer pairs used for detecting a 19p13.2 region of a human chromosome 19 and 7 groups of primer pairs used for detecting a 19q13.33 region of the human chromosome 19, 20 kinds of tag primers used for distinguishing samples, a GC stabilizer, a PCR enzyme mixture, purifying magneticbeads, a quality control standard substance and ddH2O. Through two-time PCR amplification and two-time product purification and sequencing, through a ratio of READS(1p) / READS(1q) to READS(19q) / READS(19p), the heterozygosity loss state of 1p19q in the samples is analyzed. By adopting the method, the variation situation of the 1p19q can be reflected comprehensively, truly and accurately, and gene variation of the 96 samples can be subjected to high-throughput detection and analysis simultaneously, so that the detection cost is greatly reduced.