Recombinase Polymerase Amplification Assay for Rapid Diagnostics of Dengue Infection
In: ISSN: 1932-6203, 2015
academicJournal
Zugriff:
International audience ; BACKGROUND:Over 2.5 billion people are exposed to the risk of contracting dengue fever (DF). Early diagnosis of DF helps to diminish its burden on public health. Real-time reverse transcription polymerase amplification assays (RT-PCR) are the standard method for molecular detection of the dengue virus (DENV). Real-time RT-PCR analysis is not suitable for on-site screening since mobile devices are large, expensive, and complex. In this study, two RT-recombinase polymerase amplification (RT-RPA) assays were developed to detect DENV1-4.METHODOLOGY/PRINCIPAL FINDINGS:Using two quantitative RNA molecular standards, the analytical sensitivity of a RT-RPA targeting the 3´non-translated region of DENV1-4 was found to range from 14 (DENV4) to 241 (DENV1-3) RNA molecules detected. The assay was specific and did not cross detect other Flaviviruses. The RT-RPA assay was tested in a mobile laboratory combining magnetic-bead based total nucleic acid extraction and a portable detection device in Kedougou (Senegal) and in Bangkok (Thailand). In Kedougou, the RT-RPA was operated at an ambient temperature of 38 °C with auxiliary electricity tapped from a motor vehicle and yielded a clinical sensitivity and specificity of 98% (n=31) and 100% (n=23), respectively. While in the field trial in Bangkok, the clinical sensitivity and specificity were 72% (n=90) and 100%(n=41), respectively.CONCLUSIONS/SIGNIFICANCE:During the first 5 days of infection, the developed DENV1-4 RT-RPA assays constitute a suitable accurate and rapid assay for DENV diagnosis. Moreover, the use of a portable fluorescence-reading device broadens its application potential to the point-of-care for outbreak investigations.
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Recombinase Polymerase Amplification Assay for Rapid Diagnostics of Dengue Infection
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Autor/in / Beteiligte Person: | Abd El Wahed, Ahmed ; Patel, Pranav ; Faye, Oumar ; Thaloengsok, Sasikanya ; Heidenreich, Doris ; Matangkasombut, Ponpan ; Manopwisedjaroen, Khajohnpong ; Sakuntabhai, Anavaj ; Sall, Amadou, Alpha ; Hufert, Frank ; Weidmann, Manfred ; Mansoura University Egypt ; Robert Koch Institute Berlin (RKI) ; Arbovirus et Virus de Fièvres Hémorragiques Dakar, Sénégal ; Institut Pasteur de Dakar ; Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP) ; Mahidol University Bangkok ; University Medical Center Göttingen (UMG) ; Center of Excellence for Vectors and Vector-Borne Diseases (CVVD) ; Génétique fonctionnelle des Maladies infectieuses - Functional Genetics of Infectious Diseases ; Institut Pasteur Paris (IP)-Centre National de la Recherche Scientifique (CNRS) ; Medizinische Hochschule Brandenburg Theodor Fontane / Brandenburg Medical School Theodor-Fontane (MHB Theodor Fontane) ; University of Stirling ; This study was funded by a grant from the Deutsche Gesellschaft für Internationale Zusammenarbeit (GIZ) contract no: VN 811-40270. http://www.giz.de. The Bangkok cohort study was funded by the Office of the Higher Education Commission and Mahidol University under the National Research Universities Initiative, and European Commission Seventh Framework Programme FP7/2007-2013 for the DENFREE project under Grant Agreement number 282 378. http://cordis.europa.eu/fp7/health/. The funders had no role in design of the study, data collection and analysis, decision to publish, or preparation of the manuscript. ; European Project: 282378,EC:FP7:HEALTH,FP7-HEALTH-2011-single-stage,DENFREE(2012) |
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Zeitschrift: | ISSN: 1932-6203, 2015 |
Veröffentlichung: | HAL CCSD ; Public Library of Science, 2015 |
Medientyp: | academicJournal |
DOI: | 10.1371/journal.pone.0129682 |
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