Development of a wide-range real-time RT-PCR assay for detection of Avian Reovirus (ARV)

Elsevier

Available online 7 September 2022, 114613

Journal of Virological MethodsHighlights•

Developed a specific molecular-based technique for early detection of ARV through the RT-qPCR method.

The new primers targeted for the L1 segment were tested for their sensitivity and specificity.

The RT-qPCR analysis reveals that the developed method can detect up to 1 × 101 copies/μl.

The current RT-qPCR technique has a 100-fold increase in sensitivity over the traditional RT-PCR.

This assay will provide a platform for the rapid identification of ARVs in veterinary diagnostic laboratories.

Abstract

Avian reovirus (ARV) is a common pathogen in chickens and other birds causing a variety of clinical symptoms such as arthritis and tenosynovitis but also enteric and respiratory symptoms. A rapid method that detects as many ARV genotypes as possible, will contribute to the early identification and control of the virus infection that causes high economic damage to the poultry industry worldwide. In this study, a real-time reverse transcription polymerase chain reaction (RT-qPCR) assay for the detection of ARV was developed. The RT-qPCR detection threshold for ARV genomic RNA standard cases was 10 copies/ µL. Reproducibility of the RT-qPCR was confirmed by intra- and inter-assays. When the nucleic acids of different ARV genotypes and other common avian pathogens (IBDV, AIV, NDV, and IBV) were subjected to that RT-qPCR test, only ARV samples tested positive while all other pathogens tested negative. Due to the simplicity, convenience, high sensitivity, and specificity of the assay, the probe-based RT-qPCR is proposed to be used as an alternative diagnostic assay for the detection of ARVs in veterinary diagnostic laboratories.

Keywords

Avian orthreovirus

Real-time RT-PCR

TaqMan

Sensitivity

Specificity

wide-range detection

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