Abstract

Background
In certain cases, low-level mutations that carry essential diagnostic or therapeutic signals may
be obscured by the presence of excessive, undamaged, wild-type DNA (wtDNA). Cancer,
infectious illnesses, prenatal diagnosis, organ transplantation, and prenatal diagnosis are all
examples of this problem. Since the early 1990s, approaches for mutation enrichment have
evolved that allow undesired DNA to be reduced, allowing for the discovery of low-level
alterations. They are always being improved and updated with the latest technology. New and
improved methods for practical and successful mutant enrichment and detection of low-level
mutations of therapeutic significance are increasingly in demand because of the growing
popularity of liquid biopsy for residual cancer monitoring, resistance identification, and early
diagnosis of cancer.

Content
Enzymatic techniques, PCR-blocking technologies, and physicochemical approaches are all
included in this overview of newly discovered mutant enrichment technologies. Technologies
that enable pre-PCR blockage of wtDNA to bypass PCR errors [nuclease-aided minor allele
enrichment assay with overlapping probes (NaME-PrO) and ultraviolet cross-linking minor allele
enrichment (UVME)] and provide high multiplexity followed by next-generation sequencing
[Minor allele enriched sequencing through recognition oligonucleotides (MAESTRO)] are given
particular attention.

Synopsis This paper provides an update on the state of rare mutant enrichment technology
since its inception in 2000. Traditional criteria for accuracy, speed, and cost-effectiveness all
hold true for liquid biopsy, but the field's expansion need better limits of detection. In light of
these new specifications, an assessment is made of the existing state of technology.