Mutation Enrichment Technologies Advance Low-Frequency Variant Detection for Liquid Biopsy
This review surveys twelve years of innovation in mutation enrichment technologies designed to unmask low-frequency variants hidden by excess wild-type DNA. The authors describe PCR-blocking, enzymatic, and physicochemical approaches, with particular emphasis on NaME-PrO, UVME, and the highly multiplexed MAESTRO platform for NGS-based detection. These methods address critical needs in liquid biopsy for residual cancer monitoring, resistance detection, and early cancer screening where improved limits of detection are essential.
The original study
Recent Developments in Mutation Enrichment and Detection Technologies.
- Authors
- Darbeheshti F, Yu F, Ahmed F, Adalsteinsson VA, Makrigiorgos GM
- Journal
- Clinical chemistry
- Type
- Journal Article, Review, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't
- PMID
- 35716101
Original abstract
BACKGROUND: Presence of excess unaltered, wild-type DNA (wtDNA) providing information of little clinical value may often mask low-level mutations containing important diagnostic or therapeutic clues. This is a recurring hurdle in biotechnology and medicine, including cancer, prenatal diagnosis, infectious diseases, and organ transplantation. Mutation enrichment techniques that allow reduction of unwanted DNA to enable the detection of low-level mutations have emerged since the early 1990s. They are continuously being refined and updated with new technologies. The burgeoning interest in liquid biopsies for residual cancer monitoring, detection of resistance to therapy, and early cancer detection has driven an expanded interest in new and improved methodologies for practical and effective mutation enrichment and detection of low-level mutations of clinical relevance. CONTENT: Newly developed mutation enrichment technologies are described and grouped according to the main principle of operation, PCR-blocking technologies, enzymatic methods, and physicochemical approaches. Special emphasis is given to technologies enabling pre-PCR blockage of wtDNA to bypass PCR errors [nuclease-assisted minor-allele enrichment assay with overlapping probes (NaME-PrO) and UV-mediated cross-linking minor allele enrichment (UVME)] or providing high multiplexity followed by next-generation sequencing [Minor allele enriched sequencing through recognition oligonucleotides (MAESTRO)]. SUMMARY: This review summarizes technological developments in rare mutation enrichment over the last 12 years, complementing pre-2010 reviews on this topic. The expanding field of liquid biopsy calls for improved limits of detection (LOD) and highly parallel applications, along with the traditional requirements for accuracy, speed, and cost-effectiveness. The current technologies are reviewed with regards to these new requirements.