Evaluation of SYBR Green Based Real-Time Polymerase Chain Reaction (qPCR) in Detection of Reovirus

Abstract

Avian reovirus (ARV) belongs to the Orthoreovirus genus and Reoviridae family and contains icosahedral symmetry with a double-shelled arrangement of surface protein. Avian reoviruses infection can often be seen in a broad range of avian species, including chickens, ducks, pigeons, and turkeys. These viruses can cause tenosynovitis, causing acute lameness due to swelling of one or both hock joints, and can be associated with malabsorption, hepatitis, myocarditis, respiratory disease, and enteritis. This study objective was to use the BRYT Green® based real-time RT-PCR assay in targeting the S4 segment gene for the detection of AVR, thus improving the efficiency and cost of the diagnosis. Thirty-one (n=31), including known ARV-positive samples consisting of pool organs, liver, tendon, and allantoic fluid, were selected to be tested using BRYT Green®-based real-time RT-PCR. The finding of this study showed the one-step BRYT Green®-based RT-qPCR protocol for the detection of the Reovirus S4 genome segment is a success and has advantages compared with conventional RT-PCR. This assay is time-efficient, cost-effective, user-friendly, sensitive, and highly specific. There is no doubt that this protocol can be used in many laboratory diagnoses for avian reovirus infection.