Loop-mediated isothermal amplification: Rapid and cost-effective method for detection of pathogens
Subject Areas : Diagnostic Microbiology
Ahzam Askari
1
,
Mohammad Kargar
2
,
Sadegh Ghorbani-Dalini
3
,
Abbas Doosti
4
1 - Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran.
2 - Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran.
3 - Department of Microbiology, Jahrom Branch, Young Researcher’s clube Islamic Azad University, Jahrom, Iran.
4 - Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran.
Keywords: Optimization, LAMP, Primer Design,
Abstract :
Loop-mediated isothermal amplification (LAMP) is nucleic acid amplification method that amplifies target isothermally at 62-65˚C. In this approach, withouth thermal denaturation step, double stranded DNA is simultaneousely denatured and synthesized by using the Bst (Bacillus stearothrmophilus) DNA polymerase with high strand displacement activity. Thus the reaction can be conducted with a cost-effective quipment such as water bath or heating block, and the thermal-cycling needs of a PCR are avoided. The LAMP reaction uses six primers that specifically recognize eight distinct regions on the target sequence. Moreover, the LAMP reaction produces a large amount of amplified products, resulting in easier detection, such as visual judgment based on the turbidity or colour change result from fluorescent dsDNA intercalating dye in the reaction mixture, so gel electrophoresis is not required. Therefore, the LAMP assay has the advantages of high specificity, sensitivity, amplification efficiency and simple detection, that without the need for expensive equipment could be applicable as valuable tool for rapid diagnosis of infectious diseases in both clinical and hospital laboratories of developing countries. The aim of this article is to introduce the principles and applications of LAMP method for detection of infectious agents.
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