A real-time fluorescence loop-mediated isothermal amplification method for the detection of Anisakis simplex

We established a real- time fluorescence loop- mediated isothermal amplification( LAMP) approach for the sensitive,rapid and reliable detection of Anisakis simplex.The method was based on LAMP reaction.Three sets of the specificity and sensitivity of primers were designed from ITS2 r DNA. The specificity and sensitivity were tested.Specific amplification products were obtained with A.simplex,while no amplification products were detected with DNA of related parasites,demonstrating the specificity of the assay.The capable detection of plasmid DNA in the method was 1 fg / μL.The real- time fluorescence LAMP assay was proved to be 100 times more sensitive than a conventional polymerase chain reaction for detection of A. simplex. Samples testing by the real- time fluorescence LAMP method and PCR method showed that,the result of the LAMP method was the same with PCR method.The established real- time fluorescence LAMP method was suitable for specifically detecting A.simplex.