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Background  Human schistosomiasis, mainly due to  Schistosoma mansoni  species, is one of the most prevalent parasitic diseases worldwide. To overcome the drawbacks of classical parasitological and serological methods in detecting  S. mansoni  infections, especially in acute stage of the disease, development of cost-effective, simple and rapid molecular methods is still needed for the diagnosis of schistosomiasis. A promising approach is the loop-mediated isothermal amplification (LAMP) technology. Compared to PCR-based assays, LAMP has the advantages of reaction simplicity, rapidity, specificity, cost-effectiveness and higher amplification efficiency. Additionally, as results can be inspected by the naked eye, the technique has great potential for use in low-income countries.    Methodology/Principal findings  A sequence corresponding to a mitochondrial  S. mansoni  minisatellite DNA region was selected as a target for designing a LAMP-based method to detect  S. mansoni  DNA in stool samples. We used a  S. mansoni  murine model to obtain well defined stool and sera samples from infected mice with  S. mansoni  cercariae. Samples were taken weekly from week 0 to 8 post-infection and the Kato-Katz and ELISA techniques were used for monitoring the infection. Primer set designed were tested using a commercial reaction mixture for LAMP assay and an  in house  mixture to compare results. Specificity of LAMP was tested using 16 DNA samples from different parasites, including several  Schistosoma  species, and no cross-reactions were found. The detection limit of our LAMP assay (SmMIT-LAMP) was 1 fg of  S. mansoni  DNA. When testing stool samples from infected mice the SmMIT-LAMP detected  S. mansoni  DNA as soon as 1 week post-infection.    Conclusions/Significance  We have developed, for the first time, a cost-effective, easy to perform, specific and sensitive LAMP assay for early detection of  S. mansoni  in stool samples. The method is potentially and readily adaptable for field diagnosis and disease surveillance in schistosomiasis-endemic areas.

plos neglected tropical diseases

A Loop-Mediated Isothermal Amplification (LAMP) Assay for Early Detection of Schistosoma mansoni in Stool Samples: A Diagnostic Approach in a Murine Model