A pond-side test for Guinea worm: Development of a loop-mediated isothermal amplification (LAMP) assay for detection of Dracunculus medinensis
| Autor: | Robbie A. McDonald, Sioban Ostoja-Starzewska, Neil Boonham, Jenny Tomlinson |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Time Factors 030231 tropical medicine Immunology Guinea worm Loop-mediated isothermal amplification Zoology GUINEA WORM INFECTION Biology Disease Vectors Diagnostic tools Sensitivity and Specificity Article Copepoda 03 medical and health sciences 0302 clinical medicine Dogs LAMP parasitic diseases Animals Humans Lack of knowledge Ponds Diagnostics DNA Primers Larva Base Sequence fungi General Medicine 030108 mycology & parasitology DNA Helminth Dracunculus Nematode biology.organism_classification Infectious Diseases Molecular Diagnostic Techniques Africa Cats Parasitology human activities Nucleic Acid Amplification Techniques Dracunculus medinensis Copepod Worm infection Papio |
| Zdroj: | Experimental Parasitology |
| ISSN: | 1090-2449 |
| Popis: | Guinea worm Dracunculus medinensis causes debilitating disease in people and is subject to an ongoing global eradication programme. Research and controls are constrained by a lack of diagnostic tools. We developed a specific and sensitive LAMP method for detecting D. medinensis larval DNA in copepod vectors. We were able to detect a single larva in a background of field-collected copepods. This method could form the basis of a “pond-side test” for detecting potential sources of Guinea worm infection in the environment, in copepods, including in the guts of fish as potential transport hosts, enabling research, surveillance and targeting of control measures. The key constraint on the utility of this assay as a field diagnostic, is a lack of knowledge of variation in the temporal and spatial distribution of D. medinensis larvae in copepods in water bodies in the affected areas and how best to sample copepods to obtain a reliable diagnostic sample. These fundamental knowledge gaps could readily be addressed with field collections of samples across areas experiencing a range of worm infection frequencies, coupled with field and laboratory analyses using LAMP and PCR. Highlights • LAMP tests were developed for Dracunculus medinensis and D. insignis and were shown to be specific and sensitive. • A LAMP test was developed to amplify DNA from copepods to use as an internal amplification control during testing. • Samples of copepods taken from ponds could be tested using the LAMP tests and Dracunculus medinensis could be detected. • Results are achieved in less than 30 min using just the Genie III instrument and no other laboratory equipment is necessary. |
| Databáze: | OpenAIRE |
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