Establishment of loop-mediated isothermal amplification assay for detection of parasitic ciliate Ichthyophthirius multifiliis in cyprinid fish

The objective of this research was to establish loop-mediated isothermal amplifications (LAMP) that could be used detect parasitic ciliate Ichthyophthirius multifiliis (I. multifiliis) in freshwater cyprinid fish. Primers were developed from the distinguishing fragments 18S ribosomal RNA I. and LAMP test then evaluate optimize various concentrations chemicals, time temperature. results indicated required 1.6 ?M FIP primers BIP primers, 0.2 F3 B3, 2 mM Mg2+, 1 M Betaine, 0.6 dNTP. This assay able parasite DNA within a 40 min period incubation at constant optimal temperature 64oC. positive sample appeared as clear ladder like pattern on gel electrophoresis, while yellowish green color with SYBR Green I under ultraviolet light use heating block. determined more sensitive than conventional PCR detection multifiliis. In conclusion, we have presented specific rapid system for based amplification. Importantly, employed an alternative effective diagnostic method place other molecular techniques.