Among various isothermal amplification systems developed over the recent years, the most frequently applied approach seems to be loop-mediated isothermal amplification (LAMP), implemented first by Notomi. Briefly, each reaction is carried out four oligonucleotide primers which recognize six distinct regions on the target DNA in conjunction with two loop primers to accelerate the reaction. LAMP assay can also amplify nucleic acid under isothermal condition in the range of 60 to 65ðC, all turbidity- and fluorescent-based detections, as well as agarose gel electrophoresis system are applied to visualize suspicious samples. LAMP-positive amplicons confirmed by adding a number of fluorescent dsDNA intercalating dye including ethidium bromide, SYBR Green I and propidium iodide after the reaction is completed or metal indicators such as calcein , GeneFinderââ¢, hydroxynaphthol blue (HNB) and magnesium pyrophosphate prior to the reaction, allowing observation with the naked eye. LAMP assay was widely applied for detection of many important infectious diseases in human and animals.
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Citation: Almasi MA, Dehabadi SMH, Moradi A, Eftekhari Z, Ojaghkandi MA, et al. (2013) Development and Application of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Fusarium Oxysporum f. Sp. lycopersici. J Plant Pathol Microb 4:177
Last date updated on September, 2024