Detecting Fasciola hepatica and F. gigantica microRNAs using loop-mediated isothermal amplification (LAMP)

Abstract

   Fascioliasis is a parasitic infection typically caused by two common parasites of class Trematodo, genus Fasciola, named Fasciola hepatica and F. gigantica. The widespread appearance of these species in water and food makes fascioliasis become a global zoonotic disease that affects 2.4 million people in more than 75 countries worldwide. Typically, F. hepatica and F. gigantica can be recognized with parasitological techniques to detect Fasciola spp. eggs, immunological techniques to detect worm-specific antibodies, or molecular techniques for instance polymerase chain reactions to detect parasitic genomic DNA. Recently, miRNAs have been recognised a key regulator and potential diagnostic biomarkers of diseases, including parasitic infection. An isothermal PCR called LAMP (loop-mediated isothermal amplification) is rapid, sensitive, and this amplification is very extensive, making it well-suited for field diagnostics. LAMP reaction for miRNA detection has been introduced and is able to detect miRNA in the range between 1.0amol and 1.0pmol, showing high selectivity to differentiate one miRNA sequence from others. Here, we introduced a modified LAMP to detect a typical miRNA of both F. hepatica and F. gigantica. Our method does not demand an initial heating step and the reactions have a high sensitivity even 1,000 times higher in comparison to that reported in previous studies. These results create a promising technique basis for some novel and simple device to diagnose fascioliasis and other parasitic diseases at point-of-care.