Molecular typing of N gene and 3′ untranslated region of IBV field isolates and vaccine strains using RT-PCR and RFLP

• Published in: Comparative clinical pathology Vol. 21; no. 3; pp. 283 - 287
• Main Authors: Majdani, Raheleh, Mardani, Karim, Morshedi, Ahmad, Marandi, Mehdi Vasfi, Talebi, Alireza, Yazdani, Iraj
• Format: Journal Article
• Language: English
• Published: London Springer-Verlag 01.06.2012; Springer Nature B.V
• Subjects: Hematology; Infectious bronchitis virus; Medicine; Medicine & Public Health; Oncology; Original Article; Pathology; Polymerase chain reaction; Restriction fragment length polymorphism; 3′ Untranslated region; Nucleocapsid gene; Infectious bronchitis virus
• ISSN: 1618-5641; 1618-565X
• DOI: 10.1007/s00580-010-1093-3

Whole nucleocapsid (N) gene and 3′ untranslated 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, Alu I and Mnl I. 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 3′ UTR, 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 3′ UTR 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.