Exploration of isothermal nucleic acid amplification techniques in the biomedical field

Sukanya Kundu , Rajat Varshney , Sourabh Sulabh
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Abstract

Nucleic acid amplification techniques (NATs) are frequently used in the molecular biology arena. Polymerase chain reaction (PCR) and its variants are one of the most popular NATs. The requirement of a sophisticated thermocycler and skilled technician for PCR limits its use in resource-limited laboratories and fields. Alternatively, the isothermal amplification technique can also deliver proficiency, simplicity, sensitivity, and fidelity without the need for the thermocycler. Several isothermal methods have been devised and newer concepts are also emerging. Some of them are loop-mediated isothermal amplification, whole genome amplification, rolling circle amplification, nucleic acid sequence-based amplification, and polymerase spiral reaction. These cost-effective, practicable, and easy-to-perform diagnostic assays are in current trends to identify pathogens, tumors, embryo sex, and genetically modified organisms. Isothermal amplification can also be used in microfluidic devices and point-of-need diagnostics. This review focuses on the development of common isothermal processes, their characteristics, and their acceptance over the PCR.

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探索生物医学领域的等温核酸扩增技术
核酸扩增技术(NAT)是分子生物学领域经常使用的技术。聚合酶链反应(PCR)及其变体是最常用的 NATs 之一。PCR 需要精密的热循环仪和熟练的技术人员,这限制了它在资源有限的实验室和领域的使用。另一种方法是等温扩增技术,无需热循环仪,也能提供熟练、简单、灵敏和保真度。目前已设计出多种等温方法,更新的概念也在不断涌现。其中包括环介导等温扩增、全基因组扩增、滚动圈扩增、基于核酸序列的扩增和聚合酶螺旋反应。这些诊断方法成本低、实用性强、操作简便,是目前鉴定病原体、肿瘤、胚胎性别和转基因生物的趋势。等温扩增还可用于微流控设备和需求点诊断。本综述将重点介绍常见等温扩增过程的发展、特点以及相对于 PCR 的接受程度。
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