Einzeltreffer — DigiBib

International audience ; BACKGROUND:4 one-step, real-time, reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays were developed for the detection of dengue virus (DENV) serotypes by considering 2,056 full genome DENV sequences. DENV1 and DENV2 RT-LAMP assays were validated with 31 blood and 11 serum samples from Tanzania, Senegal, Sudan and Mauritania. DENV3 and DENV4 RT-LAMP assays were validated with 25 serum samples from Cambodia.METHODOLOGY/PRINCIPAL FINDINGS:4 final reaction primer mixes were obtained by using a combination of Principal Component Analysis of the full DENV genome sequences, and LAMP primer design based on sequence alignments using the LAVA software. These mixes contained 14 (DENV1), 12 (DENV2), 8 (DENV3) and 3 (DENV4) LAMP primer sets. The assays were evaluated with an External Quality Assessment panel from Quality Control for Molecular Diagnostics. The assays were serotype-specific and did not cross-detect with other flaviviruses. The limits of detection, with 95% probability, were 22 (DENV1), 542 (DENV2), 197 (DENV3) and 641 (DENV4) RNA molecules, and 100% reproducibility in the assays was obtained with up to 102 (DENV1) and 103 RNA molecules (DENV2, DENV3 and DENV4). Validation of the DENV2 assay with blood samples from Tanzania resulted in 23 samples detected by RT-LAMP, demonstrating that the assay is 100% specific and 95.8% sensitive (positive predictive value of 100% and a negative predictive value of 85.7%). All serum samples from Senegal, Sudan and Mauritania were detected and 3 untyped as DENV1. The sensitivity of RT-LAMP for DENV4 samples from Cambodia did not quite match qRT-PCR.CONCLUSIONS/SIGNIFICANCE:We have shown a novel approach to design LAMP primers that makes use of fast growing sequence databases. The DENV1 and DENV2 assays were validated with viral RNA extracted clinical samples, showing very good performance parameters.

Titel:	Development and validation of four one-step real-time RT-LAMP assays for specific detection of each dengue virus serotype.
Autor/in / Beteiligte Person:	Lopez-Jimena, Benjamin ; Bekaert, Michaël ; Bakheit, Mohammed ; Frischmann, Sieghard ; Patel, Pranav ; Simon-Loriere, Etienne ; Lambrechts, Louis ; Duong, Veasna ; Dussart, Philippe ; Harold, Graham ; Fall, Cheikh ; Faye, Oumar ; Sall, Amadou, Alpha ; Weidmann, Manfred ; Institute of Aquaculture Stirling ; University of Stirling ; MAST Diagnostica GmbH Reinfeld ; Robert Koch Institute Berlin (RKI) ; Génétique fonctionnelle des Maladies infectieuses - Functional Genetics of Infectious Diseases ; Institut Pasteur Paris (IP)-Centre National de la Recherche Scientifique (CNRS) ; Interactions Virus-Insectes - Insect-Virus Interactions (IVI) ; Institut Pasteur du Cambodge ; Réseau International des Instituts Pasteur (RIIP) ; Arbovirus et Virus de Fièvres Hémorragiques Dakar, Sénégal ; Institut Pasteur de Dakar ; Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP) ; Institute of Aquaculture ; This study has been funded by the EU research project DiscoGnosis (Disc-shaped Point-of-Care platform for infectious disease diagnosis). This project was funded by the European Commission under the 7th Framework Programme 2007-2013 for Research and Technological Development of the EU (Grant Agreement No. 318408) (BLJ, MBa, SF, MW). The authors acknowledge the support of the MASTS pooling initiative (The Marine Alliance for Science and Technology for Scotland) in the completion of this study. MASTS is funded by the Scottish Funding Council (grant reference HR09011) and contributing institutions (MBe). ; European Project: 318408,EC:FP7:ICT,FP7-ICT-2011-8,DISCOGNOSIS(2012)
Link:	https://pasteur.hal.science/pasteur-01973492 https://pasteur.hal.science/pasteur-01973492/document Volltext https://doi.org/10.1371/journal.pntd.0006381
Zeitschrift:	ISSN: 1935-2727, 2018
Veröffentlichung:	HAL CCSD ; Public Library of Science, 2018
Medientyp:	academicJournal
DOI:	10.1371/journal.pntd.0006381
Schlagwort:	MESH: DNA Primers/genetics MESH: Dengue/virology MESH: Nucleic Acid Amplification Techniques/instrumentation MESH: Nucleic Acid Amplification Techniques/methods MESH: Phylogeny MESH: RNA Viral/genetics MESH: Reverse Transcription MESH: Tanzania MESH: Dengue Virus/classification MESH: Dengue Virus/genetics MESH: Dengue Virus/isolation & purification MESH: Dengue Virus/metabolism MESH: Humans [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology
Sonstiges:	Nachgewiesen in: BASE Sprachen: English Collection: Réseau International des Instituts Pasteur, Paris: HAL-RIIP Document Type: article in journal/newspaper Language: English Relation: info:eu-repo/semantics/altIdentifier/pmid/29813062; info:eu-repo/grantAgreement/EC/FP7/318408/EU/Disc-shaped point-of-care platform for infectious disease diagnosis/DISCOGNOSIS; pasteur-01973492; https://pasteur.hal.science/pasteur-01973492; https://pasteur.hal.science/pasteur-01973492/document; https://pasteur.hal.science/pasteur-01973492/file/pntd.0006381.pdf; PUBMED: 29813062; PUBMEDCENTRAL: PMC5973574 Rights: http://creativecommons.org/licenses/by/ ; info:eu-repo/semantics/OpenAccess

Klicken Sie ein Format an und speichern Sie dann die Daten oder geben Sie eine Empfänger-Adresse ein und lassen Sie sich per Email zusenden.

BibTeX Citavi, JabRef, u.a.
(Literaturverwaltung)

PDF kein Volltext!
(Merkzettel, Notizen)

RIS Endnote, Citavi u.a.
(Literaturverwaltung)

MODS
(XML zur Weiterverarbeitung)

oder

Wählen Sie das für Sie passende Zitationsformat und kopieren Sie es dann in die Zwischenablage, lassen es sich per Mail zusenden oder speichern es als PDF-Datei.

Gewünschter Zitations-Stil:

oder

Bitte prüfen Sie, ob die Zitation formal korrekt ist, bevor Sie sie in einer Arbeit verwenden. Benutzen Sie gegebenenfalls den "Exportieren"-Dialog, wenn Sie ein Literaturverwaltungsprogramm verwenden und die Zitat-Angaben selbst formatieren wollen.

Development and validation of four one-step real-time RT-LAMP assays for specific detection of each dengue virus serotype.