Sie befinden Sich nicht im Netzwerk der Universität Paderborn. Der Zugriff auf elektronische Ressourcen ist gegebenenfalls nur via VPN oder Shibboleth (DFN-AAI) möglich. mehr Informationen...
Rapid detection and subtyping of European swine influenza viruses in porcine clinical samples by haemagglutinin‐ and neuraminidase‐specific tetra‐ and triplex real‐time RT ‐ PCR s
Ist Teil von
Influenza and other respiratory viruses, 2016-11, Vol.10 (6), p.504-517
Ort / Verlag
Chichester: John Wiley & Sons, Inc
Erscheinungsjahr
2016
Link zum Volltext
Quelle
Wiley Online Library - AutoHoldings Journals
Beschreibungen/Notizen
Background
A diversifying pool of mammalian‐adapted influenza A viruses (
IAV
) with largely unknown zoonotic potential is maintained in domestic swine populations worldwide. The most recent human influenza pandemic in 2009 was caused by a virus with genes originating from
IAV
isolated from swine. Swine influenza viruses (
SIV
) are widespread in European domestic pig populations and evolve dynamically. Knowledge regarding occurrence, spread and evolution of potentially zoonotic
SIV
in Europe is poorly understood.
Objectives
Efficient
SIV
surveillance programmes depend on sensitive and specific diagnostic methods which allow for cost‐effective large‐scale analysis.
Methods
New
SIV
haemagglutinin (
HA
) and neuraminidase (
NA
) subtype‐ and lineage‐specific multiplex real‐time
RT
‐
PCR
s (
RT
‐
qPCR
) have been developed and validated with reference virus isolates and clinical samples.
Results
A diagnostic algorithm is proposed for the combined detection in clinical samples and subtyping of
SIV
strains currently circulating in Europe that is based on a generic, M‐gene‐specific influenza A virus
RT
‐
qPCR
. In a second step, positive samples are examined by tetraplex
HA
‐ and triplex
NA
‐specific
RT
‐
qPCR
s to differentiate the porcine subtypes H1, H3, N1 and N2. Within the
HA
subtype H1, lineages “av” (European avian‐derived), “hu” (European human‐derived) and “pdm” (human pandemic A/H1N1, 2009) are distinguished by
RT
‐
qPCR
s, and within the
NA
subtype N1, lineage “pdm” is differentiated. An
RT
‐
PCR
amplicon Sanger sequencing method of small fragments of the
HA
and
NA
genes is also proposed to safeguard against failure of multiplex
RT
‐
qPCR
subtyping.
Conclusions
These new multiplex
RT
‐
qPCR
assays provide adequate tools for sustained
SIV
monitoring programmes in Europe.