Single molecule targeted sequencing for cancer gene mutation detection

Yan Gao, Liwei Deng, QinYan, Yongqian Gao, Zengding Wu, Jinsen Cai, Daorui Ji, Gailing Li, Ping Wu, Huan Jin, Luyang Zhao, Song Liu, Liangjin Ge, Michael W. Deem & Jiankui He


With the rapid decline in cost of sequencing, it is now affordable to examine multiple genes in a single disease-targeted clinical test using next generation sequencing. Current targeted sequencing methods require a separate step of targeted capture enrichment during sample preparation before sequencing. Although there are fast sample preparation methods available in market, the library preparation process is still relatively complicated for physicians to use routinely. Here, we introduced an amplifcation-free Single Molecule Targeted Sequencing (SMTS) technology, which combined targeted capture and sequencing in one step. We demonstrated that this technology can detect low-frequency mutations using artifcially synthesized DNA sample. SMTS has several potential advantages, including simple sample preparation thus no biases and errors are introduced by PCR reaction. SMTS has the potential to be an easy and quick sequencing technology for clinical diagnosis such as cancer gene mutation detection, infectious disease detection, inherited condition screening and noninvasive prenatal diagnosis.