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

Published online by Cambridge University Press:  09 March 2017

KOSALA G. WEERAKOON ,
CATHERINE A. GORDON ,
PENGFEI CAI ,
GEOFFREY N. GOBERT ,
MARY DUKE ,
GAIL M. WILLIAMS  and
DONALD P. MCMANUS
KOSALA G. WEERAKOON*
Affiliation:
Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Brisbane, Australia School of Public Health, University of Queensland, Brisbane, Australia Department of Parasitology, Faculty of Medicine and Allied Sciences, Rajarata University of Sri Lanka, Saliyapura, Sri Lanka
CATHERINE A. GORDON
Affiliation:
Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Brisbane, Australia
PENGFEI CAI
Affiliation:
Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Brisbane, Australia
GEOFFREY N. GOBERT
Affiliation:
Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Brisbane, Australia
MARY DUKE
Affiliation:
Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Brisbane, Australia
GAIL M. WILLIAMS
Affiliation:
School of Public Health, University of Queensland, Brisbane, Australia
DONALD P. MCMANUS
Affiliation:
Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Brisbane, Australia
*
*Corresponding author: Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Brisbane, Australia. E-mail: [email protected]
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Summary

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

SchistosomiasisSchistosoma japonicumdroplet digital PCRmouse modeldiagnosis
Type
Research Article
Information
Parasitology , Volume 144 , Issue 8 , July 2017 , pp. 1005 - 1015
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Copyright
Copyright © Cambridge University Press 2017 

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Footnotes

†

Present address: Queen's University Belfast, School of Biological Sciences, Belfast, UK.

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