Abstract
This review describes the application of peptide nucleic acids (PNAs) as clamps that prevent nucleic acid amplification of wild-type DNA so that DNA with mutations may be observed. These methods are useful to detect single-nucleotide polymorphisms (SNPs) in cases where there is a small amount of mutated DNA relative to the amount of normal (unmutated/wild-type) DNA. Detecting SNPs arising from mutated DNA can be useful to diagnose various genetic diseases, and is especially important in cancer diagnostics for early detection, proper diagnosis, and monitoring of disease progression. Most examples use PNA clamps to inhibit PCR amplification of wild-type DNA to identify the presence of mutated DNA associated with various types of cancer.
Highlights
Single nucleotide changes occurring within a normal DNA sequence may be associated with different diseases, and in particular, the development or progression of various cancers [1]
We report on the use of peptide nucleic acids (PNA)-based clamps with a specific emphasis on their application to the identification of various cancers
Nagai et al [75] developed a detection system for epidermal growth factor receptor (EGFR) mutations using a combination of PNA clamps to suppress amplification of wild-type DNA and locked nucleic acids (LNA) with fluorescent groups as probes to signal the presence of mutant DNA
Summary
Single nucleotide changes occurring within a normal (often called wild-type) DNA sequence may be associated with different diseases, and in particular, the development or progression of various cancers [1]. Some existing methods are polymerase chain reaction (PCR) restriction fragment length polymorphism mapping (PCR-RFLP), allele-specific PCR (AS-PCR), allele-specific hydrolysis or dual hybridization probes, high resolution melting analysis (HRMA), amplification refractory mutation system (ARMS), dual priming oligonucleotides (DPO), TaqMan allelic discrimination assay, pyrosequencing, generation sequencing (NGS), IntPlex, BEAMing, and droplet digital PCR (dPCR) [26,27,28,29,30,31,32,33,34,35] Most of these methods can detect DNA with a single mutation when they are present at only 1% to 5% relative to the amount of wild-type DNA in a sample. We report on the use of peptide nucleic acids (PNA)-based clamps with a specific emphasis on their application to the identification of various cancers
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