The invention discloses a Y-chromosome STR gene locus fluorescence labelled multiplex amplification kit with reinforcing resolution capability. The kit comprises the following specific oligonucleotides amplification primer pairs of 27 Y-chromosome STR gene loci: DYS456, DYS458, DYS437, DYS439, DYS392, DYS385a/b, DYS393, DYS391, DYS390, DYS635, DYS438, DYS389I, DYS19, GATA-H4, DYS389II and DYS448, wherein the kit is characterized by additionally comprising at least one of the specific oligonucleotides amplification primer pairs of 10 Y-chromosome STR gene loci: DYS576, DYS570, DYS481, DYS627, DYS460, DYS449, DYS533, DYS518 and DYF387S1. The number of the detection loci of the kit is greater than that of the market similar product both at home and abroad, so that the discrimination power and the combined power of exclusion of the system are greatly improved and the individual discriminability is generally improved. Simultaneously the invention creatively comprises 7 rapid mutation sites, so that the discrimination capacity of the close relative males can be increased, the practicability and the functionality of the kit are greatly increased, and a vicious cycle of blindly increasing the similar detection sites by the domestic kit developers is avoided.