Please use this identifier to cite or link to this item: http://hdl.handle.net/1893/22809
Appears in Collections:Aquaculture Journal Articles
Peer Review Status: Refereed
Title: Development of reverse transcription recombinase polymerase amplification assay for avian influenza H5N1 HA gene detection
Authors: Yehia, Nahed
Arafa, Abdel-Satar
El, Wahed Ahmed Abd
El-Sanousi, Ahmed A
Weidmann, Manfred
Shalaby, Mohamed A
Contact Email: m.w.weidmann@stir.ac.uk
Keywords: Avian influenza
Subtype H5N1
Recombinase polymerase amplification assay
Real-time RT-PCR
Issue Date: Oct-2015
Publisher: Elsevier
Citation: Yehia N, Arafa A, El Wahed AA, El-Sanousi AA, Weidmann M & Shalaby MA (2015) Development of reverse transcription recombinase polymerase amplification assay for avian influenza H5N1 HA gene detection, Journal of Virological Methods, 223, pp. 45-49.
Abstract: The 2006 outbreaks of H5N1 avian influenza in Egypt interrupted poultry production and caused staggering economic damage. In addition, H5N1 avian influenza viruses represent a significant threat to public health. Therefore, the rapid detection of H5 viruses is very important in order to control the disease. In this study, a qualitative reverse transcription recombinase polymerase amplification (RT-RPA) assay for the detection of hemagglutinin gene of H5 subtype influenza viruses was developed. The results were compared to the real-time reverse transcription polymerase chain reaction (RT-PCR). Anin vitrotranscribed RNA standard of 970 nucleotides of the hemagglutinin gene was developed and used to determine the assay sensitivity. The developed H5 RT-RPA assay was able to detect one RNA molecule within 7min, while in real-time RT-PCR, at least 90min was required. H5 RT-RPA assay did not detect nucleic acid extracted from H5 negative samples or from other pathogens producing respiratory manifestation in poultry. The clinical performance of the H5 RT-RPA assay was tested in 30 samples collected between 2014 and 2015; the sensitivity of H5 RT-RPA and real-time RT-PCR was 100%. In conclusion, H5 RT-RPA was faster than real-time RT-PCR and easily operable in a portable device. Moreover, it had an equivalent sensitivity and specificity.
Type: Journal Article
URI: http://hdl.handle.net/1893/22809
DOI Link: http://dx.doi.org/10.1016/j.jviromet.2015.07.011
Rights: The publisher does not allow this work to be made publicly available in this Repository. Please use the Request a Copy feature at the foot of the Repository record to request a copy directly from the author. You can only request a copy if you wish to use this work for your own research or private study.
Affiliation: National Laboratory for Veterinary Quality Control on Poultry Production
National Laboratory for Veterinary Quality Control on Poultry Production
German Primate Center
Cairo University
Aquaculture
Cairo University

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