Logo des Robert Koch-InstitutLogo des Robert Koch-Institut
Publikationsserver des Robert Koch-Institutsedoc
de|en
Publikation anzeigen 
  • edoc Startseite
  • Artikel in Fachzeitschriften
  • Artikel in Fachzeitschriften
  • Publikation anzeigen
  • edoc Startseite
  • Artikel in Fachzeitschriften
  • Artikel in Fachzeitschriften
  • Publikation anzeigen
JavaScript is disabled for your browser. Some features of this site may not work without it.
Gesamter edoc-ServerBereiche & SammlungenTitelAutorSchlagwortDiese SammlungTitelAutorSchlagwort
PublizierenEinloggenRegistrierenHilfe
StatistikNutzungsstatistik
Gesamter edoc-ServerBereiche & SammlungenTitelAutorSchlagwortDiese SammlungTitelAutorSchlagwort
PublizierenEinloggenRegistrierenHilfe
StatistikNutzungsstatistik
Publikation anzeigen 
  • edoc Startseite
  • Artikel in Fachzeitschriften
  • Artikel in Fachzeitschriften
  • Publikation anzeigen
  • edoc Startseite
  • Artikel in Fachzeitschriften
  • Artikel in Fachzeitschriften
  • Publikation anzeigen
2013-02-14Zeitschriftenartikel DOI: 10.1186/1743-422X-10-58
Development of one-step quantitative reverse transcription PCR for the rapid detection of flaviviruses
Patel, Pranav
Landt, Olfert
Kaiser, Marco
Faye, Oumar
Koppe, Tanja
Lass, Ulrich
Sall, Amadou A.
Niedrig, Matthias
Background: The genus Flavivirus includes several pathogenic agents that cause severe illness in humans. Re-emergence of West Nile virus in Europe and continuous spread of certain flaviviruses such as dengue, yellow fever and Japanese encephalitis viruses represent a global danger to public health. Therefore, a rapid and accurate molecular method is required for diagnostics and epidemiological surveillance of flaviviruses. Methods: A Pan-Flavi quantitative RT-PCR assay using a Locked-Nucleic Acid probe targeting the flavivirus NS5 gene was developed and optimized to detect a wide range of flaviviruses simultaneously. The specificity and sensitivity of the Pan-Flavi assay were tested using RNA of different flaviviruses and non-flaviviruses. Furthermore, the assay was compared directly to flavivirus species-specific assays for the ability to detect flaviviruses sensitively. Results: Two degenerate primers and one Locked-Nucleic Acids probe were designed to amplify most of the flaviviruses. To increase the specificity and fluorescence signal of the Pan-Flavi assay for detection of yellow fever virus and dengue virus 4, additional primers and probes were included. Viral RNA of thirty different flaviviruses was detected, verifying the broad range specificity. The testing of this assay was successful, using standard plasmid and RNA dilutions of yellow fever virus vaccine strain, dengue virus 1 and tick-borne encephalitis virus, with a sensitivity limit of 10–100 genome copies/reaction. Also comparatively good results were achieved for detecting different flaviviruses by the Pan-Flavi assay when compared to the flavivirus species-specific assays. Conclusion: The assay is rapid, broad-range flavivirus-specific and highly sensitive making it a valuable tool for rapid detection of flaviviruses in livestock samples, epidemiological studies or as useful complement to single flavivirus-specific assays for clinical diagnosis.
Dateien zu dieser Publikation
Thumbnail
275IdTrQEYJk.pdf — PDF — 1.340 Mb
MD5: e9e58a68d628df9b03a468e9b94ec183
Zitieren
BibTeX
EndNote
RIS
Keine Lizenzangabe
Zur Langanzeige
Nutzungsbedingungen Impressum Leitlinien Datenschutzerklärung Kontakt

Das Robert Koch-Institut ist ein Bundesinstitut im

Geschäftsbereich des Bundesministeriums für Gesundheit

© Robert Koch Institut

Alle Rechte vorbehalten, soweit nicht ausdrücklich anders vermerkt.

 
DOI
10.1186/1743-422X-10-58
Permanent URL
https://doi.org/10.1186/1743-422X-10-58
HTML
<a href="https://doi.org/10.1186/1743-422X-10-58">https://doi.org/10.1186/1743-422X-10-58</a>