A pond-side test for Guinea worm: Development of a loop-mediated isothermal amplification (LAMP) assay for detection of Dracunculus medinensis

Exp Parasitol. 2020 Oct:217:107960. doi: 10.1016/j.exppara.2020.107960. Epub 2020 Aug 2.

Abstract

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.

Keywords: Diagnostics; Dracunculus medinensis; Guinea worm; LAMP.

MeSH terms

  • Africa
  • Animals
  • Base Sequence
  • Cats
  • Copepoda / genetics
  • Copepoda / parasitology*
  • DNA Primers / chemistry
  • DNA, Helminth / isolation & purification
  • Disease Vectors
  • Dogs
  • Dracunculus Nematode / genetics
  • Dracunculus Nematode / isolation & purification*
  • Humans
  • Molecular Diagnostic Techniques / standards*
  • Nucleic Acid Amplification Techniques / standards*
  • Papio
  • Ponds / parasitology*
  • Sensitivity and Specificity
  • Time Factors

Substances

  • DNA Primers
  • DNA, Helminth

Supplementary concepts

  • LAMP assay