Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2017 Jun 21;14(1):120.
doi: 10.1186/s12985-017-0788-3.

Chorioallantoic membranes of embryonated chicken eggs as an alternative system for isolation of equine influenza virus

Ilona Marcelina Gora  1 , Malgorzata Kwasnik  1 , Jan Franciszek Zmudzinski  1 , Wojciech Rozek  2
Affiliations

Chorioallantoic membranes of embryonated chicken eggs as an alternative system for isolation of equine influenza virus

Ilona Marcelina Gora et al. Virol J. .

Abstract

Background: Influenza virus isolation in embryonated chicken eggs (ECEs) is not applicable for rapid diagnosis, however it allows the recovery and propagation of the viable virus. A low number of infectious virus particles in the swabs, poor quality of samples or individual strain properties can lead to difficulties during the virus isolation process. We propose to utilize chorioallantoic membranes (CAM) of ECEs with the assistance of real-time RT PCR to facilitate equine influenza virus isolation.

Methods: Real-time RT PCR was used to detect influenza virus genetic material in amniotic/allantoic fluids (AF) and CAM of ECEs. Haemagglutination assay was used for AF. We used highly diluted virus as a substitute of clinical specimen for ECEs inoculation.

Results: Our study demonstrated that real-time RT PCR testing of CAM homogenates was more useful than testing of AF for EIV detection in ECEs. Positive results from CAM allowed to select the embryos from those with haemagglutination assay (HA) - and real-time RT PCR-negative AF for further passages. Using homogenates of CAM for subsequent passages, we finally obtained HA-positive AF, which confirmed virus replication.

Conclusion: We postulate that real-time RT PCR testing of CAM homogenates and their subsequent passages may facilitate the isolation of equine influenza viruses.

Keywords: Chorioallantoic membranes; Influenza virus; Virus isolation.

PubMed Disclaimer

References

    1. Slater J, Borchers K, Chambers T, Cullinane A, Duggan V, Elton D, Legrand L, Paillot R, Fortier G. Report of the international equine influenza roundtable expert meeting at le Touquet, Normandy, February 2013. Equine Vet J. 2014;46:645–650. doi: 10.1111/evj.12302. - DOI - PubMed
    1. Galvin P, Gildea S, Nelly M, Quinlivan M, Arkins S, Walsh C, Cullinane A. The evaluation of three diagnostic tests for the detection of equine influenza nucleoprotein in nasal swabs. Influenza Other Respir Viruses. 2014;8:376–383. doi: 10.1111/irv.12235. - DOI - PMC - PubMed
    1. Oxburgh L, Hagström A. A PCR based method for the identification of equine influenza virus from clinical samples. Vet Microbiol. 1999;67:161–174. doi: 10.1016/S0378-1135(99)00041-3. - DOI - PubMed
    1. Lu Z, Chambers TM, Boliar S, Branscum AJ, Sturgill TL, Timoney PJ, Reedy SE, Tudor LR, Dubovi EJ, Vickers ML, et al. Development and evaluation of one-step TaqMan real-time reverse transcription-PCR assays targeting nucleoprotein, matrix, and hemagglutinin genes of equine influenza virus. J Clin Microbiol. 2009;47:3907–3913. doi: 10.1128/JCM.00598-09. - DOI - PMC - PubMed
    1. Quinlivan M, Dempsey E, Ryan F, Arkins S, Cullinane A. Real-time reverse transcription PCR for detection and quantitative analysis of equine influenza virus. J Clin Microbiol. 2005;43:5055–5057. doi: 10.1128/JCM.43.10.5055-5057.2005. - DOI - PMC - PubMed

Publication types

MeSH terms

LinkOut - more resources