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Title: The use of recombinant influenza proteins for the development of avian influenza virus diagnostics
Author: Phapugrankul, Pongsathon
ISNI:       0000 0004 2726 8381
Awarding Body: University of Reading
Current Institution: University of Reading
Date of Award: 2011
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The possible emergence of a highly pathogenic avian influenza virus (HPAI) which can cross the species barrier to infect new hosts, avian or mammalian, has become a serious threat and public concern. The timely diagnosis and isolation of such viruses is crucial for outbreak control and the development of drug and vaccine responses. Modelling suggests that the next outbreak is likely to occur in rural area where the practical difficulties of management would be enormous. Therefore, the development of rapid and simple diagnostic tests to identify the index case of HPAI infection, under field conditions, is essential. Currently available rapid diagnostic tests for influenza A viruses detection suffer from low sensitivity and a lack of the ability to identify potential pandemic subtypes. This study sought to develop a rapid diagnostic test for type A influenza viruses as well as investigate the potential for identification of pandemic subtypes HS, H7, and H9. The aim was to demonstrate that tests could be developed in-house that were more sensitive, cost- effective, and easy to use than the existing commercial products, notably by using immunomagnetic beads. To do this, antigens and polyclonal antibodies necessary for using in the detection of influenza A viruses as well as potential pandemic subtype HS, H7, and H9 infections were prepared starting from the sequence analysis through to the purification process. The material generated was shown to be highly selective for their target analytes and did not cross-react with antigens or antibodies from other related respiratory diseases. In particular, the NP protein showed an equivalent performance to the existing commercial rapid test products: ClVTEST (HIPRA, Spain) and FlockChek (lDEXX, USA), in identifying type A influenza virus antibodies. Moreover, when fabricated with magnetic bead particles, an NP-immunomagnetic bead assay was shown to improve the analytical sensitivity in detecting influenza A virus protein ~320 times when compared with a conventional ELlSA. Although subtype specifi c assays for HS, H7, and H9 remain challenging due to their highly variable antigenic structure, immunomagnetic bead assays succeeded in demonstrating an improvement in not only the analytical sensitivity but also the specificity of the H7 HA subtype diagnosis when compared with a standard ELlSA. Therefore, immunomagnetic bead assay have the potential to be further developed and used as rapid test to improve the sensitivity of influenza virus diagnosis under field conditions.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available