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Rapid molecular detection of Zika virus in urine using the recombinase polymerase amplification assay

Ahmed Abd El Wahed, Sabri S. Sanabani, Oumar Faye, Rodrigo Pessôa, João Veras Patriota, Ricardo Rodrigues Giorgi, Pranav Patel, Susanne Böhlken-Fascher, Olfert Landt, Matthias Niedrig, de A. Zanotto Paolo M., Claus-Peter Czerny, Amadou A. Sall, Manfred Weidmann
doi: https://doi.org/10.1101/078501
Ahmed Abd El Wahed
1Division of Microbiology and Animal Hygiene, Department of Animal Sciences, Georg-August-University, Goettingen, Germany
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  • For correspondence: abdelwahed@gwdg.de abdelwahed@me.com
Sabri S. Sanabani
2Laboratory of Medical Investigation in Dermatology and Immunodeficiency, LIM-56/03. São Paulo Institute of Tropical Medicine, Faculty of Medicine, University of São Paulo, São Paulo, Brazil
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Oumar Faye
3Arbovirus and viral hemorragic fever unit, Institut Pasteur de Dakar, Dakar, Senegal
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Rodrigo Pessôa
2Laboratory of Medical Investigation in Dermatology and Immunodeficiency, LIM-56/03. São Paulo Institute of Tropical Medicine, Faculty of Medicine, University of São Paulo, São Paulo, Brazil
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João Veras Patriota
4Municipal Hospital of Tuparetama, Pernambuco, Brazil
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Ricardo Rodrigues Giorgi
5University of Santo Amaro, São Paulo, Brazil
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Pranav Patel
6TIB MOLBIOL Syntheselabor GmbH, Berlin, Germany
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Susanne Böhlken-Fascher
1Division of Microbiology and Animal Hygiene, Department of Animal Sciences, Georg-August-University, Goettingen, Germany
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Olfert Landt
6TIB MOLBIOL Syntheselabor GmbH, Berlin, Germany
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Matthias Niedrig
7Robert Koch Institute, Berlin, Germany
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de A. Zanotto Paolo M.
8Laboratory of Molecular Evolution and Bioinformatics, Department of Microbiology, Biomedical Sciences Institute, University of São Paulo, São Paulo, Brazil
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Claus-Peter Czerny
1Division of Microbiology and Animal Hygiene, Department of Animal Sciences, Georg-August-University, Goettingen, Germany
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Amadou A. Sall
3Arbovirus and viral hemorragic fever unit, Institut Pasteur de Dakar, Dakar, Senegal
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Manfred Weidmann
9Institute of Aquaculture, University of Stirling, Stirling, Scotland, United Kingdom
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Abstract

Background Currently the detection of Zika virus (ZIKV) in patient samples is done by real-time RT-PCR. Samples collected from rural area are sent to highly equipped laboratories for screening. A rapid point-of-care test is needed to detect the virus, especially at low resource settings.

Methodology/Principal Findings In this report, we describe the development of a reverse transcription isothermal recombinase polymerase amplification (RT-RPA) assay for the identification of ZIKV. RT-RPA assay was portable, sensitive (21 RNA molecules), and rapid (3-15 minutes). No cross-reactivity was detected to other flaviviruses, alphaviruses and arboviruses. Compared to real-time RT-PCR, the diagnostic sensitivity was 92%, while the specificity was 100%.

Conclusions/Significance The developed assay is a promising platform for rapid point of need detection of ZIKV in low resource settings and elsewhere (e.g. during mass gathering).

Author Summary Currently, Dengue (DENV), Zika (ZIKV) and Chikungunya (CHIKV) viruses represent a global threat. The clinical picture of the acute febrile diseases caused by DENV, ZIKV and CHIKV is very similar, in addition, the same mosquito vector is involved in the transmission cycle. The differentiation between them is of great importance as supportive treatment differs and the identification of any of the three viruses prompts implementation of control measures to avoid spreading of an outbreak. We have developed an assay for the detection of ZIKV genome. The assay based on isothermal “recombinase polymerase amplification” assay, which was performed at one temperature (42°C). The result was obtained in maximum of 15 minutes. Moreover, the assay is easy to be implemented at low resource settings.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted October 03, 2016.
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Rapid molecular detection of Zika virus in urine using the recombinase polymerase amplification assay
Ahmed Abd El Wahed, Sabri S. Sanabani, Oumar Faye, Rodrigo Pessôa, João Veras Patriota, Ricardo Rodrigues Giorgi, Pranav Patel, Susanne Böhlken-Fascher, Olfert Landt, Matthias Niedrig, de A. Zanotto Paolo M., Claus-Peter Czerny, Amadou A. Sall, Manfred Weidmann
bioRxiv 078501; doi: https://doi.org/10.1101/078501
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Rapid molecular detection of Zika virus in urine using the recombinase polymerase amplification assay
Ahmed Abd El Wahed, Sabri S. Sanabani, Oumar Faye, Rodrigo Pessôa, João Veras Patriota, Ricardo Rodrigues Giorgi, Pranav Patel, Susanne Böhlken-Fascher, Olfert Landt, Matthias Niedrig, de A. Zanotto Paolo M., Claus-Peter Czerny, Amadou A. Sall, Manfred Weidmann
bioRxiv 078501; doi: https://doi.org/10.1101/078501

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