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Several technologies for detection and quantitation of sequence variations have been developed for genomics research and molecular diagnostics, namely polymerase chain reaction (PCR), isothermal amplification, hybridization, and next-generation sequencing (NGS). NGS allows the massively multiplexed sequence analysis of DNA and RNA and, thus, it is optimal method for multiplexed analysis of many genes and their variants. However, NGS has a significant error rate due to signal ambiguity, enzyme infidelity, imperfect deprotection and others, making the method very inefficient in case of the low frequency targets. PCR is more accurate than microarrays or NGS, it has high molecular sensitivity, and ease of use. Most of the FDA approved assays for molecular diagnostics are based on PCR methods. The main weakness of PCR is the primer dimer formation that result in false positives or false negatives. The …