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

Yigal Farnoushi; Dan Heller; Avishai Lublin

doi:10.1016/j.jviromet.2022.114613

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

J Virol Methods. 2022 Dec:310:114613. doi: 10.1016/j.jviromet.2022.114613. Epub 2022 Sep 7.

Authors

Yigal Farnoushi¹, Dan Heller², Avishai Lublin³

Affiliations

¹ Department of Avian Diseases, Kimron Veterinary Institute, Beit Dagan 5025001, Israel; Koret School of Veterinary Medicine, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot 7610001, Israel. Electronic address: igalfa@moag.gov.il.
² Koret School of Veterinary Medicine, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot 7610001, Israel.
³ Department of Avian Diseases, Kimron Veterinary Institute, Beit Dagan 5025001, Israel.

PMID: 36087792
DOI: 10.1016/j.jviromet.2022.114613

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 orthoreovirus; Real-time RT-PCR; Sensitivity; Specificity; TaqMan; Wide-range detection.

MeSH terms

Animals
Chickens
Nucleic Acids*
Orthoreovirus, Avian* / genetics
Poultry Diseases* / diagnosis
RNA
Reoviridae Infections* / diagnosis
Reoviridae Infections* / veterinary
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction
Sensitivity and Specificity

Substances

RNA
Nucleic Acids