Specific and quantitative assay for duck variant orthoreovirus utilizing the loop-mediated isothermal amplification technique

Abstract

Since 2018 and up to the present, newly discovered avian orthoreovirus (NDRV) types have been a prominent cause of illness concerns in ducks in China. Infected birds with the NDRV frequently had significant splenic necrosis. For effective avian reovirus (ARV) infection management and prevention, it is essential to have access to improved diagnostic techniques for quickly diagnosing recently discovered ARV variations. To identify NDRVs from clinical samples of various avian origins, a unique quantitative loop-mediated isothermal amplification assay (qLAMP) was created in this work. By utilizing 10-fold serial dilutions of the standard substance, excellent low detection limits (1.0 copies/μL) were observed. The test had no cross-reaction with avian viruses other than NDRV, and the generated qLAMP could successfully identify NDRV reference strains. Intra- and interassay tests with variability ranging from 0.08% to 2.78% were used to demonstrate the assay's reproducibility. The qLAMP results demonstrated a greater detection efficiency when compared to those from the examination of 90 clinical samples using traditional PCR and RT-qPCR. Overall, this assay was sensitive and specific for the identification of NDRV clinical samples from diverse bird species and should be used as a standard diagnostic laboratory test.