Isothermal Amplification of Nucleic Acids: The Race for the Next “Gold Standard”

Beatriz Oliveira, B. Veigas, P. Baptista
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引用次数: 55

Abstract

Nucleic acid amplification technologies (NAATs) have become fundamental tools in molecular diagnostics, due to their ability to detect small amounts of target molecules. Since its development, Polymerase Chain Reaction (PCR) has been the most exploited method, being stablished as the “gold standard” technique for DNA amplification. However, the requirement for different working temperatures leads to the need of a thermocycler machine or complex thermal apparatus, which have been preventing its application in novel integrated devices for single workflow and high throughput analysis. Conversely, isothermal amplification methods have been gaining attention, especially for point-of-care diagnosis and applications. These non-PCR based methods have been developed by mimicking the in vivo amplification mechanisms, while performing the amplification with high sensitivity, selectivity and allowing for high-throughput analysis. These favorable capabilities have pushed forward the implementation and commercialization of several platforms that exploit isothermal amplification methods, mostly against virus, bacteria and other pathogens in water, food, environmental and clinical samples. Nevertheless, the future of isothermal amplification methods is still dependent on achieving technical maturity and broader commercialization of enzymes and reagents.
核酸等温扩增:下一个“金标准”的竞赛
核酸扩增技术(NAAT)由于能够检测少量目标分子,已成为分子诊断的基本工具。自发展以来,聚合酶链式反应(PCR)一直是最受欢迎的方法,被确立为DNA扩增的“金标准”技术。然而,对不同工作温度的要求导致了对热循环机或复杂热设备的需求,这阻碍了其在用于单一工作流程和高通量分析的新型集成设备中的应用。相反,等温扩增方法越来越受到关注,尤其是在护理点诊断和应用方面。这些基于非PCR的方法是通过模拟体内扩增机制开发的,同时以高灵敏度、高选择性进行扩增,并允许高通量分析。这些有利的能力推动了几个利用等温扩增方法的平台的实施和商业化,这些方法主要针对水、食品、环境和临床样本中的病毒、细菌和其他病原体。然而,等温扩增方法的未来仍然取决于酶和试剂的技术成熟度和更广泛的商业化。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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