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Characterization And Phylogenetic Analysis Of Hemagglutinin Gene Of Avian Influenza Virus Subtype H9n2 Isolated In 2015

By: Arslan Mehboob (2009-VA-76) | Prof. Dr. Tahir Yaqub.
Contributor(s): Dr. Jawad Nazir | Dr. Maryam Javed.
Material type: materialTypeLabelBookPublisher: 2016Description: 43p.Subject(s): Department of MicrobiologyDDC classification: 2474-T Dissertation note: H9N2 Avian influenza outbreaks has caused great economic losses to poultry industry resulting in decrease egg production, high morbidity and mortality. Due to different antigenic variants H9 has become problematic. It has the ability to cross species barrier and increase in pathogenicity. Hemagglutination inhibition (HI) test is employed extensively for subtyping and detection of antibody titre against the virus. Continuous mutations in the HA gene transforms AIV subtype H9N2 into more pathogenic virus that may have pandemic potential and can cross species barrier. Thus, it was necessary to identify various antigenic variants of H9 virus. It was important to study the HA gene as it plays vital role in viral attachment, release of genetic material and pathogenicity. In present study, a sum of four H9 virus samples were isolated. Both serological and molecular confirmation was done. 200 samples from different areas were collected and properly labelled. They were then processed for egg inoculation in embryonated eggs. Virus was grown in embryonated eggs and harvested fluid is then proceeded for confirmatory testing. Haemagllutination and Haemagllutination Inhibition testing was done. RNA was extracted by Kit method and cDNA was synthesized. Reverse Transcriptase (RT-PCR) was performed using specific primer sets and then the amplicon were run on agarose gel. The bands obtained was sent for sequencing and Phylogenetic analysis was obtained using software and tree was constructed. Protein analysis was also performed. The present study enabled us to characterize and construct Phylogenetic tree of HA gene of currently prevailing H9N2 Avian Influenza isolates in Pakistan.
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Item type Current location Collection Call number Status Date due Barcode Item holds
Thesis Thesis UVAS Library
Thesis Section
Veterinary Science 2474-T (Browse shelf) Available 2474-T
Total holds: 0

H9N2 Avian influenza outbreaks has caused great economic losses to poultry industry resulting
in decrease egg production, high morbidity and mortality. Due to different antigenic variants H9
has become problematic. It has the ability to cross species barrier and increase in pathogenicity.
Hemagglutination inhibition (HI) test is employed extensively for subtyping and detection of
antibody titre against the virus. Continuous mutations in the HA gene transforms AIV subtype
H9N2 into more pathogenic virus that may have pandemic potential and can cross species
barrier. Thus, it was necessary to identify various antigenic variants of H9 virus. It was important to
study the HA gene as it plays vital role in viral attachment, release of genetic material and
pathogenicity.
In present study, a sum of four H9 virus samples were isolated. Both serological and
molecular confirmation was done. 200 samples from different areas were collected and properly
labelled. They were then processed for egg inoculation in embryonated eggs. Virus was grown in
embryonated eggs and harvested fluid is then proceeded for confirmatory testing.
Haemagllutination and Haemagllutination Inhibition testing was done. RNA was extracted by
Kit method and cDNA was synthesized. Reverse Transcriptase (RT-PCR) was performed using
specific primer sets and then the amplicon were run on agarose gel. The bands obtained was sent
for sequencing and Phylogenetic analysis was obtained using software and tree was constructed.
Protein analysis was also performed.
The present study enabled us to characterize and construct Phylogenetic tree of HA gene of
currently prevailing H9N2 Avian Influenza isolates in Pakistan.

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