Development of Loop-Mediated Isothermal Amplification Methods for Detection of the Enteric Myxosporeans Causing Myxosporean Emaciation Disease

Abstract

In this study, we developed loop-mediated isothermal amplification (LAMP) methods for detecting the enteric myxosporeans Enteromyxum leei and Sphaerospora fugu which cause myxosporean emaciation disease. The LAMP primer sets were designed based on the sequences of the small subunit ribosomal RNA gene (SSU rDNA) of E. leei and S. fugu. Reaction time and temperature were optimized for 60 minutes at 62°C, respectively. Cross-reactivity with other myxosporeans was not observed. These LAMP methods showed detection sensitivities of 100-1,000 times higher than those of the PCR methods applied so far. These LAMP methods established in this study are the specific and sensitive methods, allowing for the rapid and reliable diagnosis of this disease.