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Diagnosis of Influenza Viruses

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Insight into Influenza Viruses of Animals and Humans

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Abstract

Accurate and rapid diagnosis of the influenza virus infection can greatly help in its prevention and control. The success of confirmatory laboratory diagnosis is to a great extent dependent on the choice and quality of specimens, and their transport and storage conditions prior to processing in the laboratory. Samples such as tracheal and cloacal swabs, faeces, tissue samples including trachea, lungs, serum, etc., (both in avian and mammalian species) should be submitted. The isolation of the virus can be carried out in embryonated chicken eggs via amniotic cavity or allantoic cavity route, primary cell culture of chicken embryo fibroblast and Madin Darby Canine Kidney, embryonic swine kidney, primary swine kidney and swine testicle or swine lung epithelial cell lines, chicken kidney cell lines and primary human respiratory epithelial cells, followed by Haemagglutination Assay, Haemagglutination-Inhibition test and plaque assays for confirming the growth of the virus in embryonated egg and cell culture, respectively. The viral antigen can be demonstrated by employing indirect immunofluorescence, AGID, Immunoperoxidase test, various ELISAs, Haemagglutination-Inhibition/Neuraminidase-Inhibition, RT-PCR and PCR-ELISA. The Haemagglutination-Inhibition, ELISA and Single Radial Haemolysis are used for measuring the antibody titre. Various nucleic acid-based detection methods include RT-PCR, RRT-PCR, real-time and multiplex-microsphere-quantitative PCR, loop-mediated isothermal amplification, in situ hybridisation, DNA micro-array, nucleic acid sequence based amplification, nucleic acid sequencing and heteroduplex mobility assay. The RT-PCR and PCR-ELISA can also be used for subtyping of the influenza viruses. The techniques used for the diagnosis of exotic and pandemic influenza viruses should only be done in referral biosecurity level 3 (BSL-3) laboratories, having appropriate disease containment facilities, by trained and skilled personnel.

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Authors and Affiliations

  1. Department of Veterinary Microbiology, LLR University of Veterinary and Animal Sciences, Hisar, 125004, Haryana, India

    Sanjay Kapoor

  2. Division of Pathology, Indian Veterinary Research Institute, Izatnagar, Bareilly, 243122, Uttar Pradesh, India

    Kuldeep Dhama

Authors
  1. Sanjay Kapoor
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  2. Kuldeep Dhama
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Correspondence to Sanjay Kapoor .

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Kapoor, S., Dhama, K. (2014). Diagnosis of Influenza Viruses. In: Insight into Influenza Viruses of Animals and Humans. Springer, Cham. https://doi.org/10.1007/978-3-319-05512-1_10

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