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International Journal of Poultry Science

.: Home > International Journal of Poultry Science > 2009 > Volume 8 Number 5 > B.P. Shankar1, R.N. Sreenivas Gowda2, B. Pattnaik1, B.H. Manjunatha Prabhu3, B.K. Sreenivas4, M.K. Vinuthan5, D. Ranjith2 and H.K. Pradhan1

Identification and Subtyping of Avian Influenza Viruses by Reverse Transcription Polymerase Chain Reaction (RT-PCR) and Agarose Gel Electrophoresis

B.P. Shankar1, R.N. Sreenivas Gowda2, B. Pattnaik1, B.H. Manjunatha Prabhu3, B.K. Sreenivas4, M.K. Vinuthan5, D. Ranjith2 and H.K. Pradhan1
1High Security Animal Disease Laboratory, IVRI, Bhopal, India 2Veterinary College, Hebbal, Bangalore-24, Karnataka, India 3Molecular Biology Genetics Units, JNCASR, Jakkur, Bangalore, India 4IAH&VB, Hebbal, Bangalore, India 5MRDG, IISc, Bangalore, India
Abstract :

Avian Influenza (AI) is caused by type A influenza virus belonging to the family orthomyxoviridae,
which is classified into 16 HA and 9 NA subtypes based on two surface glycoprotein’s haemagglutinin (HA) and neuraminidase (NA). In the present study we did identification and HA-subtyping of avian influenza virus by reverse transcription-PCR (RT-PCR) during the first outbreaks of AI in India during 2006. The avian influenza virus is identified by RT-PCR using a set of primers specific to the nucleoprotein (NP) gene of avian influenza virus. The HA-subtypes of avian influenza virus were determined by running with HA subtype specific primers for H5, H7 and H9 RT-PCR reactions, each using a set of primers specific to one HAsubtype. A total of 10,236 tissue / cloacal swab samples, received at the HSADL from various parts of the country, were processed for isolation of AI virus in embryonated chicken eggs. Out of these, 9 samples originating from poultry in Maharashtra (Navapur and Jalgaon) and Gujarat (Surat) states of India were found positive for H5 virus by RT-PCR. All samples received from outbreaks areas were tested by using all tree subtype specific primers(H5, H7 and H9) only H5 RT-PCR reactions was give the product of expected size, and thus the HA-subtype of the virus is determined. One sample gave the positive result with H9 subtype specific primers. The RT-PCR procedure is rapid and sensitive, and could be used for the identification and HA-subtyping of avian influenza virus in organ homogenates.

Keywords :
Avian influenza, H5N1, RT-PCR, India

Date Deposited : 05 Jul 2011 13:09

Last Modified : 05 Jul 2011 13:09

Official URL: http://www.pjbs.org/ijps/ijps.htm

Volume 8, Number 5, - 2009 , ISSN 1682-8356

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