Ahmed Abd El Wahed1, Sabri Saeed Sanabani2, Oumar Faye3, Rodrigo Pessôa2, João Veras Patriota4, Ricardo Rodrigues Giorgi5, Pranav Patel6, Susanne Böhlken-Fascher7, Olfert Landt6, Matthias Niedrig8, Paolo Marinho de Andrade Zanotto9, Claus Peter Czerny1, Amadou A Sall3, Manfred Weidmann. 1. Division of Microbiology and Animal Hygiene, Institute of Veterinary Medicine, Department of Animal Sciences, Georg-August-University, Goettingen, Lower Saxony, Germany. 2. Hospital das Clínicas, School of Medicine, University of São Paulo, São Paulo, Brazil. 3. Arbovirus and viral hemorragic fever unit, Institut Pasteur de Dakar, Dakar, Senegal. 4. Municipal Hospital of Tuparetama, Tuparetama, Pernambuco, Brazil. 5. University of Santo Amaro, São Paulo, São Paulo, Brazil. 6. TIB MOLBIOL Syntheselabor GmbH, Berlin, Germany. 7. Georg-August-University Goettingen. 8. Robert Koch Institut Robert Koch Institut. 9. Laboratory of Molecular Evolution and Bioinformatics, Department of Microbiology, Biomedical Sciences Institute, University of São Paulo, São Paulo, Brazil.
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).
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).
Authors: Guilherme Calvet; Renato S Aguiar; Adriana S O Melo; Simone A Sampaio; Ivano de Filippis; Allison Fabri; Eliane S M Araujo; Patricia C de Sequeira; Marcos C L de Mendonça; Louisi de Oliveira; Diogo A Tschoeke; Carlos G Schrago; Fabiano L Thompson; Patricia Brasil; Flavia B Dos Santos; Rita M R Nogueira; Amilcar Tanuri; Ana M B de Filippis Journal: Lancet Infect Dis Date: 2016-02-18 Impact factor: 25.071
Authors: Ahmed Abd El Wahed; Pranav Patel; Oumar Faye; Sasikanya Thaloengsok; Doris Heidenreich; Ponpan Matangkasombut; Khajohnpong Manopwisedjaroen; Anavaj Sakuntabhai; Amadou A Sall; Frank T Hufert; Manfred Weidmann Journal: PLoS One Date: 2015-06-15 Impact factor: 3.240
Authors: Rodrigo Pessôa; João Veras Patriota; Maria de Lourdes de Souza; Alvina Clara Felix; Nubia Mamede; Sabri S Sanabani Journal: Medicine (Baltimore) Date: 2016-03 Impact factor: 1.889
Authors: Mariano Carossino; Yanqiu Li; Pei-Yu A Lee; Chuan-Fu Tsai; Pin-Hsing Chou; Dennis Williams; Ashley Skillman; R Frank Cook; Grayson Brown; Hsiao-Fen G Chang; Hwa-Tang T Wang; Udeni B R Balasuriya Journal: BMC Infect Dis Date: 2017-12-19 Impact factor: 3.090
Authors: Khalid Shahin; Jose Gustavo Ramirez-Paredes; Graham Harold; Benjamin Lopez-Jimena; Alexandra Adams; Manfred Weidmann Journal: PLoS One Date: 2018-02-14 Impact factor: 3.240
Authors: Amanda E Calvert; Brad J Biggerstaff; Nathan A Tanner; Molly Lauterbach; Robert S Lanciotti Journal: PLoS One Date: 2017-09-25 Impact factor: 3.240
Authors: Manfred Weidmann; Ousmane Faye; Oumar Faye; Ahmed Abd El Wahed; Pranav Patel; Christophe Batejat; Jean Claude Manugerra; Aimee Adjami; Matthias Niedrig; Frank T Hufert; Amadou A Sall Journal: J Infect Dis Date: 2018-10-05 Impact factor: 5.226