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1.
Iranian Journal of Veterinary Research. 2007; 62 (1): 69-80
in Persian | IMEMR | ID: emr-146226

ABSTRACT

Infectious bronchitis [IB] disease is one of the important respiratory diseases of poultry that causes annually large economic losses in poultry industry of Iran. The aim of this study is molecular characterization of S1 gene of Iranian infectious bronchitis viruse [IBV] belonged to 793/B serotype. The whole S1 gene of three local IBV strains in RT-PCR, showed a band above 1600 base pair [bp] in gel electrophoresis. RFLP analysis using 3 restriction enzymes, Hae3, Hind3 and Ecor1, showed 793/B serotype pattern. The S1 gene of these strains were sequenced and compared with 30 reference IBV strains. Identity Plot [IP] of nucleic acids and amino acids sequences were also designed. Moreover, nucleic acids differences in 1659 bp of S1 gene were calculated by distance method: nucleotide: Kimura 2-parameter and finally, the phylogenetic tree of S1 gene sequence strains with the highest validity in branching were designed. Three Iranian strains belonged to 793/B genotype with nucleotide differences of 5.64-6.07% to UK/793/B as a prototype of 793/B and 26.02-26.16% to H120 as a vaccinal strain. Regarding to low homology and weak cross protection between 793/B serotype and Massachusetts vaccinal strain, it would be a possible cause for failure in vaccination and outbreak of IB disease in vaccinated flock of poultry industry


Subject(s)
Molecular Structure , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis
2.
Journal of Veterinary Research. 2004; 59 (3): 259-264
in Persian | IMEMR | ID: emr-207076

ABSTRACT

Objective: detection of infectious bronchitis viruses by RT- PCR/RFLPs in poultry farms of Iran


Design: longitudinal study from 1997 to 2003


Samples: tracheas, lungs and kidneys of suspected flocks to respiratory diseases


Procedure: from 1997-2003, tissues samples including lung, trachea and kidney, had been prepared from broiler and layer flocks were submitted to avian virology laboratory of poultry diseases section in order to isolate respiratory viral diseases viruses. A total number of 50 infectious bronchitis viruses [IBV] were isolated in embryonated chicken eggs. The infected embryos displayed stunting, urate deposition in the mesonephros or death after three passages. Twelve isolates that had showed typical signs in embryos, were selected for molecular identification. Viral RNA was extracted by RNxTM plus [CinnaGen Co.] using chloroform 1 Isoamyl alcohol, Isopropanol, Ethanol and DEPC water. cDNA was prepared from extracted RNA with RT enzyme, RH primer, RT buffer, dNTP and Rnase inhibitor. For PCR reaction, buffer PCR ]OX, MgC12, S loligo5' and 3' primer, ampli Taq DNA polymerase and dNTP were added to cDNA and then PCR was conducted in thermal cycler. The PCR products were analyzed on a 1% agarous gel and Ethidurn Bromide staining. The S1 glycoprotein genes of IBV strains appeared to be above 1600 bp in size. PCR products were digested by HaeIII, EcorI and HindIII, according to the manufacture's recommendation


Results: base on RFLPs patterns and comparison with RFLP references patterns [793A3, D274, M41, H120], 8 of 12 strains showed 793/B pattern and the rests [4 of 12] showed Mass patterns in RFLPs


Clinical implications: regarding to low homology and weak cross protection between 793B serotype and vaccinal strain [Massachusetts] prevention and a controlled strategy against IB should be altered

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