A novel duplex ddPCR assay for the diagnosis of schistosomiasis japonica: proof of concept in an experimental mouse model

Parasitology. 2017 Jul;144(8):1005-1015. doi: 10.1017/S003118201700021X. Epub 2017 Mar 9.

Abstract

The current World Health Organization strategic plan targets the elimination of schistosomiasis as a public health problem by 2025 and accurate diagnostics will play a pivotal role in achieving this goal. DNA-based detection methods provide a viable alternative to some of the commonly used tests, notably microscopy and serology, for the diagnosis of schistosomiasis. The detection of parasite cell-free DNA in different clinical samples is a recent valuable advance, which provides significant benefits for accurate disease diagnosis. Here we validated a novel duplex droplet digital PCR assay for the diagnosis of Chinese (SjC) and Philippine (SjP) strains of Schistosoma japonicum infection in a mouse model. The assay proved applicable for both SjC and SjP infections and capable of detecting infection at a very early intra-mammalian stage in conveniently obtainable samples (urine and saliva) as well as in serum and feces. The target DNA copy numbers obtained in the assay showed a positive correlation with the infection burden assessed by direct traditional parasitology. The potential to detect parasite DNA in urine and saliva has important practical implications for large-scale epidemiological screening programmes in the future, particularly in terms of logistical convenience, and the assay has the potential to be a valuable additional tool for the diagnosis of schistosomiasis japonica.

Keywords: Schistosoma japonicum; Schistosomiasis; diagnosis; droplet digital PCR; mouse model.

Publication types

  • Validation Study

MeSH terms

  • Animals
  • Blood / parasitology
  • China
  • DNA Copy Number Variations*
  • DNA, Helminth / analysis*
  • Disease Models, Animal
  • Feces / parasitology
  • Female
  • Humans
  • Mice
  • Philippines
  • Polymerase Chain Reaction / methods*
  • Saliva / parasitology
  • Schistosoma japonicum / isolation & purification*
  • Schistosomiasis japonica / diagnosis*
  • Schistosomiasis japonica / parasitology
  • Sensitivity and Specificity
  • Urine / parasitology

Substances

  • DNA, Helminth