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2005•Acta Veterinaria BrnoOpen access

Using Nested-PCR for Detection of Avian Influenza Virus

S. P. Beladi, Seyed A. Ghorashi, Dina Morshedi

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Abstract

Avian influenza virus can infect all poultry species.Infection of Broilers and layers with this virus can cause heavy economic loss to poultry industry annually.In this study a RT-PCR was optimized to detect avian influenza virus, regardless of its subtype.A part of matrix gene which is conserved among avian influenza viruses was amplified.The PCR product was cloned in pTZ57R/T vector and sequenced.The sequence data confirmed the specificity of the test.The sensitivity of PCR was determined to be 3 × 10 -2 EID 50 /ml.Using internal primers, a nested-PCR was carried out and the sensitivity was increased to 3 × 10 -4 EID 50 /ml.Due to high sensitivity of the test for detection of avian influenza viruses, this technique can be used as a robust diagnostic method for detection of avian influenza virus. Avian influenza virus, PCR

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Avian influenza virus can infect all poultry species.Infection of Broilers and layers with this virus can cause heavy economic loss to poultry industry annually.In this study a RT-PCR was optimized to detect avian influenza virus, regardless of its subtype.A part of matrix gene which is conserved among avian influenza viruses was amplified.The PCR product was cloned in pTZ57R/T vector and sequenced.The sequence data confirmed the specificity of the test.The sensitivity of PCR was determined to be 3 × 10 -2 EID 50 /ml.Using internal primers, a nested-PCR was carried out and the sensitivity was increased to 3 × 10 -4 EID 50 /ml.Due to high sensitivity of the test for detection of avian influenza viruses, this technique can be used as a robust diagnostic method for detection of avian influenza virus. Avian influenza virus, PCR

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Available abstract

Avian influenza virus can infect all poultry species.Infection of Broilers and layers with this virus can cause heavy economic loss to poultry industry annually.In this study a RT-PCR was optimized to detect avian influenza virus, regardless of its subtype.A part of matrix gene which is conserved among avian influenza viruses was amplified.The PCR product was cloned in pTZ57R/T vector and sequenced.The sequence data confirmed the specificity of the test.The sensitivity of PCR was determined to be 3 × 10 -2 EID 50 /ml.Using internal primers, a nested-PCR was carried out and the sensitivity was increased to 3 × 10 -4 EID 50 /ml.Due to high sensitivity of the test for detection of avian influenza viruses, this technique can be used as a robust diagnostic method for detection of avian influenza virus. Avian influenza virus, PCR

Key concepts: Czech, Virology, Influenza A virus subtype H5N1, Nested polymerase chain reaction, Veterinary medicine, Biology, Avian influenza virus, Virus

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