1. Towards a Rapid-Turnaround Low-Depth Unbiased Metagenomics Sequencing Workflow on the Illumina Platforms
- Author
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Winston Lian Chye Koh, Si En Poh, Chun Kiat Lee, Tim Hon Man Chan, Gabriel Yan, Kiat Whye Kong, Lalita Lau, Wai Yip Thomas Lee, Clark Cheng, Shawn Hoon, and Yiqi Seow
- Subjects
metagenomic sequencing ,host depletion ,DNA/RNA library preparation ,liquid biopsy ,infectious disease ,Technology ,Biology (General) ,QH301-705.5 - Abstract
Unbiased metagenomic sequencing is conceptually well-suited for first-line diagnosis as all known and unknown infectious entities can be detected, but costs, turnaround time and human background reads in complex biofluids, such as plasma, hinder widespread deployment. Separate preparations of DNA and RNA also increases costs. In this study, we developed a rapid unbiased metagenomics next-generation sequencing (mNGS) workflow with a human background depletion method (HostEL) and a combined DNA/RNA library preparation kit (AmpRE) to address this issue. We enriched and detected bacterial and fungal standards spiked in plasma at physiological levels with low-depth sequencing (
- Published
- 2023
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