Rapid detection of Galba truncatula in water sources on pasture-land using loop-mediated isothermal amplification for control of trematode infections

Parasites & Vectors

Fiona Tyson¹  Emma Davies¹  Manod Williams¹  Peter M. Brophy¹  Hefin Wyn Williams¹  Chelsea N. Davis¹  Rhys Aled Jones¹  Alex Prescott¹  David Cutress¹  Dewi Llyr Jones²  Michael T. Rose³ 

[1] Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Aberystwyth, UK;Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Aberystwyth, UK;Coleg Cambria, Llysfasi, Ruthin Road, Ruthin, Denbighshire, UK;Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Aberystwyth, UK;Tasmanian Institute of Agriculture, University of Tasmania, Sandy Bay, TAS, Australia;

关键词: Loop-mediated isothermal amplification;    Environmental DNA;    Farm management;    Trematode;    Fluke;    Galba truncatula;    Fasciola hepatica;    Calicophoron daubneyi;   

DOI  :  10.1186/s13071-020-04371-0

来源: Springer

【 摘 要 】

BackgroundFascioliasis caused by the trematodes Fasciola hepatica and F. gigantica, is a global neglected zoonotic disease estimated to cost the livestock industry over €2.5 billion annually. Farm management measures and sustainable use of anthelmintics can, in principle, effectively control trematode infection in livestock and reduce the rate of developing anthelmintic resistance. Previously, we designed an environmental DNA (eDNA) assay to identify a common trematode intermediate host, the freshwater snail Galba truncatula, in water sources to measure specific trematode infection risk areas on pasture-land. To improve this procedure, we now report a loop-mediated isothermal amplification (LAMP) assay to identify G. truncatula eDNA.MethodsA LAMP assay was designed and optimised (e.g. temperature, time duration and primer concentration) to identify G. truncatula DNA. The ability of the LAMP assay to target G. truncatula DNA was identified, and LAMP assay limit of detection was investigated in comparison to conventional PCR. In the field, 48 water samples were collected from stream, ditch and water pool habitats in four locations at two Aberystwyth University farms over a seven week period to investigate the applicability of the LAMP assay for use on eDNA samples, in comparison to conventional PCR.ResultsThe LAMP assay delivered detectable results in 30 min at 63 °C. The assay discriminated between G. truncatula DNA and non-target DNA, presenting a level of DNA detection comparable to conventional PCR. No significant difference was found between the ability of the LAMP and PCR assay to identify G. truncatula eDNA in water samples. Kappa coefficient analysis revealed a moderate level of agreement between LAMP and PCR assays.ConclusionsThis study demonstrated that the LAMP assay can detect G. truncatula eDNA in a simple and rapid manner. The LAMP assay may become a valuable tool to determine optimum pasture management for trematode parasite control.

【 授权许可】

CC BY