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Development of a lateral flow recombinase polymerase assay for the diagnosis of Schistosoma mansoni infections.

Authors
  • Poulton, Kate1
  • Webster, Bonnie2
  • 1 The London School of Hygiene and Tropical Medicine, Keppel Street, London, UK; The Natural History Museum, Cromwell Road, London, UK.
  • 2 The Natural History Museum, Cromwell Road, London, UK. Electronic address: [email protected]
Type
Published Article
Journal
Analytical Biochemistry
Publisher
Elsevier
Publication Date
Feb 07, 2018
Volume
546
Pages
65–71
Identifiers
DOI: 10.1016/j.ab.2018.01.031
PMID: 29425749
Source
Medline
Keywords
License
Unknown

Abstract

Infection with Schistosoma mansoni causes intestinal schistosomiasis, a major health problem across Africa. The accurate diagnosis of intestinal schistosomiasis is vital to inform surveillance/control programs. Diagnosis mainly relies on microscopic detection of eggs in faecal samples but many factors affect sensitivity. Molecular diagnostics are sensitive and specific but application is limited as necessary infrastructure, financial resources and skilled personnel are often lacking in endemic settings. Recombinase Polymerase Amplification (RPA) is an isothermal DNA amplification/detection technology that is practical in nearly any setting. Here we developed a RPA lateral flow (LF) assay targeting the 28S rDNA region of S. mansoni. The 28S LF-RPA assay's lower limit of detection was 10pg DNA with the lower test parameters permitting sufficient amplification being 6 min and 25°C. Optimal assay parameters were 40-45°C and 10 min with an analytical sensitivity of 102 copies of DNA. Additionally the PCRD3 lateral flow detection cassettes proved more robust and sensitive compared to the Milenia HybriDetect strips. This 28S LF-RPA assay produces quick reproducible results that are easy to interpret, require little infrastructure and is a promising PON test for the field molecular diagnosis of intestinal schistosomiasis.

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