Identification of new avian Infectious Bronchitis virus variants in Iranian poultry flocks by High Resolution Melting curve analysis


Keywords:
High Resolution Melting analysis Identification Infectious Bronchitis Virus Iranian New variant Poultry
A. MASOUDIAN
N. SHEIKHI
M. H. BOZORGMEHRI-FARD
Abstract

Avian Infectious bronchitis (IB) is a common coronavirus infection of chickens and responsible for performance reduction and increasing mortality due to subsequent respiratory, renal and/or reproductive disorders. Classification of causative agent is necessary to plan successful vaccination strategies to prevent the infection due to poor inter-strains cross-reaction. To identify dominant circulating strains in Iran, a Real-time PCR combined with 3’ Un-Translated Region (3’ UTR) High Resolution Melting (HRM) analysis designed as a rapid and reliable method for IB Virus (IBV) detection and differentiation. Samples collected from 20-suspected flocks and after PCR products, HRM curves of samples as well as 6 commercial IB live vaccines with 2 standard strains, were analyzed as references. IBV genomes detected in 11 samples while according to HRM analysis and calculating Genotype Confidence Percentage (GCP), 6 positive specimens identified as 793/B field strains and the left 5 found as new IBV variant strains. Then obtained PCR products sent for nucleotide sequencing to determine genotype relativity. All five infectious agents, related to QX-like type and indicating circulation of new variants in Iran as a probable cause of vaccination failures and consequent economical losses.

Article Details
  • Section
  • Research Articles
Downloads
Download data is not yet available.
References
Adzhar, A, Shaw, K, Britton, P, Cavanagh, D (1996) Universal Oligonucleotides for the Detection of Infectious Bronchitis Virus by the Polymerase Chain Reaction. Avian Pathol 4: 817-836.
Aghakhan, S M, Abshar, N, Rasoul Nejad Fereiduni, S, Marunesi, C, Khodashenas, M (1994) Studies on Avian Viral Infections in Iran. Arch Inst Razi 45: 1-10.
Beato, M, De Battisti, C, Terregino, C, Drago, A, Capua, I, Ortali, G (2005) Evidence of Circulation of a Chinese Strain of Infectious Bronchitis Virus (Qxibv) in Italy. Vet Rec 22: 720-720.
Bochkov, Y A, Batchenko, G V, Shcherbakova, L O, Borisov, A V, Drygin, V V (2006) Molecular Epizootiology of Avian Infectious Bronchitis in Russia. Avian Pathol 5: 379 - 393.
Capua, I, Minta, Z, Karpinska, E, Mawditt, K, Britton, P, Cavanagh, D,. (1999) Co-Circulation of Four Types of Infectious Bron- chitis Virus (793/B, 624/I, B1648 and Massachusetts). Avian Pathol 6: 587 - 592.
Cavanagh, D, Gelb, J (2008) Infectious Bronchitis In: Diseases of Poultry. 12th ed. Wiley-Blackwell Publishing Iowa: pp 117-135.
Cook, J K, Jackwood, M, Jones, R (2012) The Long View: 40 Years of Infectious Bronchitis Research. Avian Pathol 3: 239-250.
Farsang, A, Ros, C, Renstrom, L H, Baule, C, Soos, T, Belak, S (2002) Molecular Epizootiology of Infectious Bronchitis Virus in Sweden Indicating the Involvement of a Vaccine Strain. Avian Pathol 3: 229-236.
Hewson, K, Noormohammadi, A H, Devlin, J M, Mardani, K, Ignjatovic, J (2009) Rapid Detection and Non-Subjective Characterisation of Infectious Bronchitis Virus Isolates Using High-Resolution Melt Curve Analysis and a Mathematical Model. Arch Virol 4: 649-660.
Hewson, K A, Browning, G F, Devlin, J M, Ignjatovic, J, Noormohammadi,A H (2010) Application of High-Resolution Melt Curve Analysis for Classification of Infectious Bronchitis Viruses in Field Specimens. Aust Vet J 10: 408-413.
Li, L, Xue, C, Chen, F, Qin, J, Xie, Q, Bi, Y,. (2010) Isolation and Genetic Analysis Revealed No Predominant New Strains of Avian Infectious Bronchitis Virus Circulating in South China During 2004-2008. Vet Microbiol 2-4: 145-154.
Lin, J-H, Tseng, C-P, Chen, Y-J, Lin, C-Y, Chang, S-S, Wu, H-S,. (2008) Rapid Differentiation of Influenza a Virus Subtypes and Genetic Screening for Virus Variants by High-Resolution Melting Analysis. J Clin Microbiol 3: 1090-1097.
Majdani, R, Mardani, K, Morshedi, A, Vasfi Marandi, M, Talebi, A (2012). Molecular Comparison of Three Different Regions of the Genome of Infectious Bronchitis Virus Field Isolates and Vaccine Strains. Vet Res Forum.
Mardani, K, Browning, G F, Ignjatovic, J, Noormohammadi, A H (2006) Rapid Differentiation of Current Infectious Bronchitis Virus Vaccine Strains and Field Isolates in Australia. Aust Vet J 1-2: 59-62.
Seyfi Abad Shapouri, M R, Mayahi, M, Assasi, K, Charkhkar,S (2004) A Survey of the Prevalence of Infectious Bronchitis Virus Type 4/91 in Iran. Acta Vet Hung 2: 163-166.
Shoushtari, A H, Toroghi, R, Momayez, R, Pourbakhsh, S A (2008) 793/B Type, the Predominant Circulating Type of Avian Infectious Bronchitis Viruses 1999-2004 in Iran: A Retrospective Study. Arch. Razi Ins. 1: 1-5.
Williams, A K, Wang, L, Sneed, L W, Collisson, E W (1993) Analysis of a Hypervariable Region in the 3’ Non-Coding End of the Infectious Bronchitis Virus Genome. Virus Research 1:19-27.
Worthington, K J, Currie, R J W, Jones, R C (2008) A Reverse Transcriptase-Polymerase Chain Reaction Survey of Infectious Bronchitis Virus Genotypes in Western Europe from 2002 to 2006. Avian Pathol 3: 247 - 257.