Detection of avian reoviruses causing tenosynovitis in breeder flocks in Iran by reverse transcription-polymerase chain reaction (RT-PCR) and restriction enzyme fragment length polymorphism (RFLP)

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Department of Avian Diseases, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

Abstract

BACKGROUND:Avian reoviruses (ARVs) are members of the
Orthoreovirus genus; one of the 12 genera of the Reoviridae
family. The ARVs are the cause of some important diseases in
poultry such as reovirus-induced arthritis, tenosynovitis,
chronic respiratory disease, and mal-absorption syndrome.
OBJECTIVES: In this study, the presence of ARVs in the Iranian
breeder flocks was investigated through reverse transcriptionpolymerase
chain reaction (RT-PCR) and restriction enzyme
fragment length polymorphism (RFLP). METHODS: A total of
800 fecal swab samples were initially collected from breeder
flocks (older than 45 weeks of age). They were then sent to the
laboratory in containers with PBS, and after that they were
pooled and finally to 120 samples were obtained. The total RNA
extracted from the pooled fecal samples were used to amplify the
selected parts of the S1 (1023 bp) and S4 (437 bp) genes from the
ARV field isolates using RT-PCR. The positive RT-PCR
amplified products were further analyzed by RFLP using five
restriction enzymes. RESULTS: Based on the findings, 5 samples
were positive with the S1 primer and 6 samples were with the S4
one. The patterns observed after the digestion of PCR products
revealed that the isolates of this study were identical to both the
S1133 vaccine and standard strains. CONCLUSIONS: The
findings suggested that the RT-PCR/RFLP analysis might be
considered as a simple and rapid approach for the differentiation
of ARVisolates. This study was the first molecular detection of
the ARVs presence in the Iranian breeder flocks using the RTPCR
amplification of the S1 and S4 genes and RFLP analysis.

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