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Detection Of Amantadine Resistant Variants Among Avian Influenza Viruses Subtype H9n2 Isolated In Pakistan

By: Sabir Subhan (2009-VA-32) | Prof. Dr. Tahir Yaqub.
Contributor(s): Dr. Muhammad Zubair Shabbir | Dr. Yasin Tipu.
Material type: materialTypeLabelBookPublisher: 2016Description: 42p.Subject(s): Department of MicrobiologyDDC classification: 2457-T Dissertation note: Avian influenza A virus subtype H9N2 is prevalent in poultry industry of Pakistan. Amantadine is an antiviral drug which is being used as prophylactic measure to control this disease despite the occurrence of resistance against this drug. There is need to monitor the resistance of amantadine in Flu viruses. In this study, we collected 100 samples of broilers birds showing mild to severe respiratory signs. Samples were collected from different locations of Punjab, Pakistan. After initial identification via Hemagglutination test, the molecular identification and confirmation of subtype H9N2 was done by multiplex PCR. To rule out the co-infection of NDV, PCR of NDV was also done. The samples which were pure H9N2 were further processed for the screening of amantadine susceptibility. To do this, titration of viruses was done on MDCK cells in the presence and absence of amantadine at the concentration of 2 ug /ml. The results of TCID5O were compared in the presence and absence of amantadine for each isolate and isolates showing difference of 2 log 10 TCID50/0.1 ml were declared resistant to amantadine as described by Masuda et al. 2000. The results of this study revealed that the viruses circulating in the poultry industry if Pakistan are resistant to this drug as we found that out 10 isolates 4 were resistant to this drug. So, there is need to monitor the usage of this drug in poultry as human cases of H9N2 viruses have been reported and virus was of avian origin. Monitoring is necessary because amantadine is recommended in flu pandemics and this virus possesses the pandemic potential and can cross the species barrier.
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Avian influenza A virus subtype H9N2 is prevalent in poultry industry of Pakistan. Amantadine is an antiviral drug which is being used as prophylactic measure to control this disease despite the occurrence of resistance against this drug. There is need to monitor the resistance of amantadine in Flu viruses. In this study, we collected 100 samples of broilers birds showing mild to severe respiratory signs. Samples were collected from different locations of Punjab, Pakistan. After initial identification via Hemagglutination test, the molecular identification and confirmation of subtype H9N2 was done by multiplex PCR. To rule out the co-infection of NDV, PCR of NDV was also done. The samples which were pure H9N2 were further processed for the screening of amantadine susceptibility. To do this, titration of viruses was done on MDCK cells in the presence and absence of amantadine at the concentration of 2 ug /ml. The results of TCID5O were compared in the presence and absence of amantadine for each isolate and isolates showing difference of 2 log 10 TCID50/0.1 ml were declared resistant to amantadine as described by Masuda et al. 2000. The results of this study revealed that the viruses circulating in the poultry industry if Pakistan are resistant to this drug as we found that out 10 isolates 4 were resistant to this drug. So, there is need to monitor the usage of this drug in poultry as human cases of H9N2 viruses have been reported and virus was of avian origin. Monitoring is necessary because amantadine is recommended in flu pandemics and this virus possesses the pandemic potential and can cross the species barrier.

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