Isothermal DNA amplification techniques are simple, rapid, cost-effective and they have the equivalent specificity and sensitivity to PCR, thus enabling point-of-care diagnostics for pathogen detection, single nucleotide polymorphism analysis, biomarker detection, etc. Future work in this area should focus on microfluidic adaptations of these techniques with integrated sample preparation and further reduction of cost and result turnover time. Effective amplification of large genomic segments by isothermal methods results in unbiased whole genome amplification from a few homogeneous cells, serving as high-quality single-cell genetic material. This material can then be used for archiving and downstream analysis such as genotyping, comparative genomic hybridization, and single-cell genomics. Immunoassays coupled with isothermal DNA amplification have improved sensitivities compared with traditional enzyme-amplified immunoassays, and improved dynamic range and reproducibility compared to immuno-PCR.
Highly specific isothermal amplification reactions (LAMP, HDA) have allowed the development of assays in which the nonspecific detection of amplicon accumulation is sufficient to indicate the presence of an initial template, allowing real-time fluorescent detection with an intercalating DNA dye such as EvaGreen or SYBR Green I.
KeywordsIsothermal NAAT Molecular diagnosis NAAT PCR
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