Development and Validation of the 9‐Dye Fluorescent Autosomal and Y Chromosome Multiplex STR Amplification System

To develop a 9-dye fluorescent autosomal and Y chromosome STR multiplex amplification system and verify its performance. Using the 9-dye fluorescent labeling technology, construct a multiplex amplification system that uses the autosomal and Y chromosome core and preferred loci recommended by GB/T 41009-2021 national standard as candidate loci, then evaluate its accuracy, sensitivity, species specificity, and other system performance index. A 9-dye fluorescent multiplex amplification system containing 70 loci, including Amel, D5S818, D21S11, D7S820, CS1FPO, D2S1338, D19S433, D8S1132, D2S441, VWA, D8S1179, D16S539, D10S1435, PentaD, D3S3045, TPOX, TH01, D22S1045, D18S51, FGA, D19S253, D10S1248, D6S477, D6S1043, D13S317, D12S391, D15S659, PentaE, D1S1656, D3S1358, DYS460, DYS389I, DYS390, DYS533, DYS392, DYS518, DYS557, DYS570, DYS458, DYS437, DYS385, GATA-H4, DYS576, DYS643, DYS508, DYS627, DYS456, DYS391, DYF404S1, DYS438, DYS448, DYF387S1, DYS596, DYS549, DYS399S1, DYS393, DYS635, DYS439, DYS19, DYS444, DYS449, DYS481, DYS527a/b, and DYS447 was successfully constructed. The various loci in this system are well balanced, with a sensitivity of 0.0625 ng and a certain ability to detect mixture samples. This system can obtain rich locus information through a single test and meet the needs of routine testing cases and DNA database construction. It has a good practical significance in forensic medicine.