Evaluation of the RTyper Y27 chip amplification system with Quick TargSeq 1.0 for Y-STR genotyping in forensic science.

The male-specific Y chromosome is widely used in forensic DNA analysis. High-mutation Y-chromosomal short tandem repeats (Y-STRs) increase the ability to differentiate related males, enhancing discrimination capacity. Current Y-STR typing requires trained forensic personnel, specialized instruments, and a controlled lab environment, taking 6-8 h. The Quick TargSeq 1.0 system integrates DNA extraction, PCR amplification, and electrophoresis via microfluidic biochip technology. Our previous study showed that Quick TargSeq can automatically produce auto-STR and DIP-38 profiles from buccal swabs, blood stains, or other biological samples in about 2 h. Here, we developed a Y-STR kit for Quick TargSeq. The RTyper Y27 microfluidic amplification system is a five-dye multiplex setup containing 27 core Y-STR loci (DYS19, DYS385a/b, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, DYS481, DYS533, DYS576, DYS635, Y-GATA-H4, DYF387S1a/b, DYS449, DYS460, DYS518, DYS627, and DYS570). To confirm the system's efficiency and accuracy, we conducted sensitivity, concordance, reproducibility, inhibition, mixture, and versatility studies following SWGDAM developmental validation guidelines. Results demonstrate that the RTyper Y27 system is efficient, accurate, and reliable for forensic applications.