The discovery of genetic factors behind increasing number of human diseases and the growth of education of genetic knowledge to the public make demands for genetic testing increase rapidly. However, traditional genetic testing methods cannot meet all kinds of the requirements.
Next generation sequencing (NGS) featured with high throughput and low cost of sequencing capacity develops fast, especially with the improvement of its read length, read accuracy and the immergence of small-sized machines, making it a powerful genetic testing tool.
In this study, we applied NGS to develop novel genetic testing methods, focused on the detection of point mutation and copy number variation (CNV). We developed a short-read multiplex sequencing method for reliable, cost-effective and high-throughput genotyping method and demonstrated its ability in a HLA genotyping test for 96 samples. In addition, we developed a targeted sequencing based preimplantation genetic diagnosis (PGD) method for monogenic diseases and tested it in a family suffering from β-thalassaemia major undergoing PGD. Moreover, we developed a method which can achieve detection of point mutation and copy number variation simultaneously by targeted sequencing and validated its accuracy in 15 samples with known results.
These novel methods, named as RCHSBT, PGD by targeted sequencing and ICLU by SeTRs respectively, were compared to their corresponding traditional and other existed NGS methods individually in terms of accuracy, simplicity, cost, throughput and turnaround time. New methods proved their superiority than current methods available, also their limitations were mentioned. In conclusion, these new methods will provide benefits to people in need.