Rapid detection of Galba truncatula in water sources on pasture-land using loop-mediated isothermal amplification for control of trematode infections
| Autor: | Emma Davies, Fiona Tyson, Michael T. Rose, David Cutress, Hefin Wyn Williams, Alex Prescott, Dewi Llyr Jones, Peter M. Brophy, Chelsea N. Davis, Manod Williams, Rhys Jones |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Fascioliasis Veterinary medicine Livestock Snails 030231 tropical medicine Loop-mediated isothermal amplification Fresh Water Environmental DNA lcsh:Infectious and parasitic diseases 03 medical and health sciences 0302 clinical medicine Animals Fasciola hepatica lcsh:RC109-216 Ecosystem Galba truncatula biology Research Farm management Calicophoron daubneyi Intermediate host food and beverages 030108 mycology & parasitology biology.organism_classification DNA Environmental Infectious Diseases Molecular Diagnostic Techniques Parasitology Primer (molecular biology) Fluke Nucleic Acid Amplification Techniques Trematode |
| Zdroj: | Parasites & Vectors Parasites & Vectors, Vol 13, Iss 1, Pp 1-11 (2020) |
| ISSN: | 1756-3305 |
| Popis: | Background Fascioliasis 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. Methods A 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. Results The 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. Conclusions This 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. |
| Databáze: | OpenAIRE |
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