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Background  Animal trypanosomosis caused by  Trypanosoma evansi  is known as “surra” and is a widespread neglected tropical disease affecting wild and domestic animals mainly in South America, the Middle East, North Africa and Asia. An essential necessity for  T. evansi  infection control is the availability of reliable and sensitive diagnostic tools. While DNA-based PCR detection techniques meet these criteria, most of them require well-trained and experienced users as well as a laboratory environment allowing correct protocol execution. As an alternative, we developed a recombinase polymerase amplification (RPA) test for Type A  T. evansi . The technology uses an isothermal nucleic acid amplification approach that is simple, fast, cost-effective and is suitable for use in minimally equipped laboratories and even field settings.    Methodology/Principle findings  An RPA assay targeting the  T. evansi  RoTat1.2 VSG gene was designed for the DNA-based detection of  T. evansi . Comparing post-amplification visualization by agarose gel electrophoresis and a lateral flow (LF) format reveals that the latter displays a higher sensitivity. The RPA-LF assay is specific for RoTat1.2-expressing strains of  T. evansi  as it does not detect the genomic DNA of other trypanosomatids. Finally, experimental mouse infection trials demonstrate that the  T. evansi  specific RPA-LF can be employed as a test-of-cure tool.    Conclusions/Significance  Compared to other DNA-based parasite detection methods (such as PCR and LAMP), the  T. evansi  RPA-LF ( Tev RPA-LF) described in this paper is an interesting alternative because of its simple read-out (user-friendly), short execution time (15 minutes), experimental sensitivity of 100 fg purified genomic  T. evansi  DNA, and ability to be carried out at a moderate, constant temperature (39°C). Therefore, the  Tev RPA-LF is an interesting tool for the detection of active  T. evansi  infections.

Development of a recombinase polymerase amplification lateral flow assay for the detection of active Trypanosoma evansi infections