Single molecule targeted sequencing for cancer gene mutation detection.

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

doi:10.1038/srep26110

Abstract

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 amplification-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 artificially 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.

Highlights

In the past few years, the cost of large-scale DNA sequencing has been dramatically driven down by the tremendous advances in next-generation sequencing (NGS)[1]
Single Molecule Targeted Sequencing (SMTS) can be applied in cancer gene mutation detection, inherited condition screening, and high-resolution human leukocyte antigen (HLA)
The fundamental limitation of detection of single molecule fluorescence signals stems from the intrinsic qualities of the fluorophore

Summary

Introduction

In the past few years, the cost of large-scale DNA sequencing has been dramatically driven down by the tremendous advances in next-generation sequencing (NGS)[1]. NGS is used to examine specific gene panels such as cancer genes and inherited conditions, sample numbers are high and data volume per sample is relatively small. It is often more cost-effective and time-efficient to target, capture, and sequence only the genomic regions of interest[2]. There are several cancer gene panels commercially available, targeting as few as 50 to many hundreds of genes that are frequently mutated in cancer patients[3]. The cancer gene panel targeted sequencing has been proved to be useful in hereditary cancers diagnosis, and disease management

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