Int. J. Environ. Res. Public Health 2014, 11(5), 4953-4964; doi:10.3390/ijerph110504953
Article

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

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Received: 21 February 2014; in revised form: 8 April 2014 / Accepted: 30 April 2014 / Published: 8 May 2014
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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
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MDPI and ACS Style

Ahmed, A.; van der Linden, H.; Hartskeerl, R.A. Development of a Recombinase Polymerase Amplification Assay for the Detection of Pathogenic Leptospira. Int. J. Environ. Res. Public Health 2014, 11, 4953-4964.

AMA Style

Ahmed A, van der Linden H, Hartskeerl RA. Development of a Recombinase Polymerase Amplification Assay for the Detection of Pathogenic Leptospira. International Journal of Environmental Research and Public Health. 2014; 11(5):4953-4964.

Chicago/Turabian Style

Ahmed, Ahmed; van der Linden, Hans; Hartskeerl, Rudy A. 2014. "Development of a Recombinase Polymerase Amplification Assay for the Detection of Pathogenic Leptospira." Int. J. Environ. Res. Public Health 11, no. 5: 4953-4964.


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