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Gewählte Publikation:

Publikationstyp: Zeitschriftenaufsatz
Dokumenttyp: Originalarbeit

Jahr: 2012

AutorInnen: Fereidouni, SR; Harder, TC; Gaidet, N; Ziller, M; Hoffmann, B; Hammoumi, S; Globig, A; Starick, E

Titel: Saving resources: avian influenza surveillance using pooled swab samples and reduced reaction volumes in real-time RT-PCR.

Quelle: J Virol Methods. 2012; 186(1-2):119-125



Autor/innen der Vetmeduni Vienna:

Fereidouni Sasan

Diese Publikation wurde nicht im Namen der Vetmeduni Vienna erstellt und ist deshalb ausschließlich der persönlichen Publikationsliste des/der Autors/Autorin zugeordnet!


Abstract:
The occurrence of highly pathogenic (HP) avian influenza (AI) H5N1 in Asia and its spread to Africa and Europe prompted costly monitoring programs of wild birds and domestic poultry. AI virus excretion is tested by examining avian swab samples by real-time reverse transcription PCR (RT-qPCR). In this study, pools of swab samples and a reagents volume reduction per RT-qPCR were evaluated as measures of economization. Viral transport medium and faecal matrices were spiked with different low pathogenic AI virus strains and tested for loss of target RNA during all processing steps as individual rayon swabs or in sample pools of 5, 10 and 15 swabs. Fresh faeces from Mallard ducks and other aquatic bird species as sample matrix resulted in loss of AIV RNA of about 90% compared to transport medium. Due to sample RNA dilution in pools the likelihood of detection of single positive samples is decreasing with increasing size of sample pools. However, pools of five samples containing only one positive sample consistently gave positive results. Similarly, no differences in detection rates were obtained when analyzing 1030 wild bird swab samples either individually or in pools of five. Reducing the reaction volume of influenza A virus generic as well as of subtype-specific RT-qPCRs to 12.5 μl (2.5 μl template) instead of 25 μl did not adversely affect the limit of detection of these RT-qPCRs. A significant economic benefit without impeding detection efficacy can be achieved when sample pools of five samples are analyzed by RT-qPCR using a reduction of the reaction mix to the half of the original volume.

Keywords Pubmed: Africa
Animals
Asia
Birds
Costs and Cost Analysis
Europe
Influenza A Virus, H5N1 Subtype/isolation & purification*
Influenza in Birds/diagnosis*
Influenza in Birds/virology
Poultry
Real-Time Polymerase Chain Reaction/economics
Real-Time Polymerase Chain Reaction/methods*
Reverse Transcriptase Polymerase Chain Reaction/economics
Reverse Transcriptase Polymerase Chain Reaction/methods*
Sensitivity and Specificity
Specimen Handling/economics
Specimen Handling/methods*
Virology/methods*


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