环介导恒温核酸扩增法在病原微生物快速检测领域的应用

Application of Loop-mediated Isothermal Amplification on Pathogens Rapid Detection Key words:

摘要:

环介导恒温核酸扩增法（loop-mediated isothermal amplification of DNA，简称LAMP）是一种新型的核酸扩增方法，可以针对靶基因的目标区域形成多种片段的扩增产物。目前对于LAMP产物的检测主要有沉淀法，荧光法和电泳法。与PCR相比，LAMP不需要模板的热变性、长时间的温度循环、繁琐的电泳等，其扩增与检测过程可以实现一步完成。因此LAMP技术具有快速、简单、高特异性、高灵敏度和成本低廉的优点。随着LAMP技术的不断完善，在可预见的将来，其将在核酸扩增领域逐渐取代PCR反应，推动检测技术向更快捷简便、更精确廉价的方向发展。目前LAMP技术已应用于致病微生物和胚胎性别鉴定等检测，其中致病微生物包括致病细菌、真菌、病毒及寄生虫等。该检测技术在食品安全、临床医疗及水产等主要领域也受到广泛应用。本文主要介绍LAMP反应的特点及其在细菌、病毒、寄生虫和水产动物病原微生物快速检测领域中的应用。

Abstract:

Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method with many fragment amplification products according to specific region of target genes. Currently, precipitation, electrophoresis and fluorescence are the main methods to detect amplification products. Compared with PCR, amplification and detection process can be achieved in one step without template thermal denaturation, long time cycle and complex electrophoresis. Therefore, LAMP has been reported and applied in the field of bacteriological detection with advantages of rapid, easy operating, low-cost, highly sensitive and specific detection. In future, it is convinced that LAMP will replace PCR assays, contributing to more rapid, simple, specific and accurate detection method. Nowadays, LAMP has been applied on detection of various pathogenic microbes and embryo sexing, and the microbes include pathogenic bacterial, fungi, virus and parasites. This detection has been employed in different fields, such as food safety, clinical medicine and fishery. This paper described the features of LAMP and its application on rapid detection of bacterial, viruses, parasites and aquatic animal pathogens.