Molecular typing of N gene and 3Πuntranslated region of IBV field isolates and vaccine strains using RT-PCR and RFLP
- نوع فایل : کتاب
- زبان : انگلیسی
- مؤلف : Raheleh Majdani & Karim Mardani & Ahmad Morshedi & Mehdi Vasfi Marandi & Alireza Talebi & Iraj Yazdani
- چاپ و سال / کشور: 2010
Description
Whole nucleocapsid (N) gene and 3Πuntranslated region (UTR) of infectious bronchitis virus (IBV) vaccines and Iranian field isolates were amplified using reverse transcription and polymerase chain reaction (RT-PCR). The amplified fragments were subjected to digestion using two restriction endonuclease enzymes, AluI and MnlI. Five different restriction fragment length polymorphism (RFLP) patterns were generated using both enzymes, classifying IBV strains into five similar groups. Based on RT-PCR and RFLP analysis of the N gene and 3ΠUTR, IBV vaccine strains H52, H120 and MA5 all from the same serotype generated identical patterns using both enzymes. Vaccine strain and field isolate of 4/91 also had the same pattern distinct from other IBVs. The IB88 vaccine strain and two other field isolates MNS-7862-1 and Ur1/09 had three different RFLP patterns distinguishable from each other and the other IBVs. In conclusion, RT-PCR and RFLP analysis of the N gene and 3ΠUTR could be employed as a useful method for differentiating IBV strains especially in cases where S1 gene amplification is not successful because of its highly variable nature among different IBVs.
Comp Clin Pathol DOI 10.1007/s00580-010-1093-3 Received: 9 April 2010 / Accepted: 25 August 2010
