Development of a Recombinase Polymerase Amplification Assay for the Detection of Pathogenic Leptospira

Ahmed Ahmed, Hans Van der Linden and Rudy A. Hartskeerl
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Ahmed Ahmed: KIT Biomedical Research, WHO/FAO/OIE and National Leptospirosis Reference Centre, Royal Tropical Institute, Meibergdreef 39, Amsterdam 1105, The Netherlands
Hans Van der Linden: KIT Biomedical Research, WHO/FAO/OIE and National Leptospirosis Reference Centre, Royal Tropical Institute, Meibergdreef 39, Amsterdam 1105, The Netherlands
Rudy A. Hartskeerl: KIT Biomedical Research, WHO/FAO/OIE and National Leptospirosis Reference Centre, Royal Tropical Institute, Meibergdreef 39, Amsterdam 1105, The Netherlands

IJERPH, 2014, vol. 11, issue 5, 1-12

Abstract: Detection of leptospires based on DNA amplification techniques is essential for the early diagnosis of leptospirosis when anti- Leptospira antibodies are below the detection limit of most serological tests. In middle and low income countries where leptospirosis is endemic, routine implementation of real-time PCR is financially and technically challenging due to the requirement of expensive thermocycler equipment. In this study we report the development and evaluation of a novel isothermal recombinase polymerase amplification assay (RPA) for detection of pathogenic Leptospira based on TwistAmp chemistry. RPA enabled the detection of less than two genome copies per reaction. Retrospective evaluation revealed a high diagnostic accuracy (sensitivity and specificity of 94.7% and 97.7%, respectively) compared to culturing as the reference standard. RPA presents a powerful tool for the early diagnosis of leptospirosis in humans and in animals. Furthermore, it enables the detection of the causative agent in reservoirs and environment, and as such is a valuable adjunct to current tools for surveillance and early outbreak warning.

Keywords: recombinase; polymerase; diagnosis; amplification; pathogenic; Leptospira; development; diagnostic; evaluation; RPA (search for similar items in EconPapers)
JEL-codes: I I1 I3 Q Q5 (search for similar items in EconPapers)
Date: 2014
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