Development and Application of a Dual RT-qPCR Assay for Detection of Classical and Novel Goose Reovirus
YIN Zhenghao
WEI Wuque
LIANG Zhaoping
LI Yuze
XU Hang
LU Yongkun
MEI Kun
HUANG Shujian
Abstract：The Goose reovirus(GRV)comprises two distinct types:the classic(GRV)and the novel goose reovirus(NGRV).These two types exhibit differences in their epidemiological characteristics and viral gene sequences.The lack of sensitive and specific assays for both types of viruses has resulted in difficulties in accurately diagnosing and timely intervening in cases.In order to distinguish GRV from NGRV,a dual RT-qPCR assay based on a dye-based method(SYBR Green I)was established.The results demonstrated that the method was capable of distinguishing GRV(79.5℃)and NGRV(86.5℃)based on their respective melting curves.Furthermore,no amplification was observed for any of the common waterfowl epidemics,the lowest detections were 6.466×101 copies/μL GRV and 4.979×101 copies/μL NGRV.The results demonstrated that the dual RT-qPCR assay,based on a dye-based method,could rapidly and accurately identify different types of GRVs in infected geese.The assay exhibited a high degree of precision,with coefficients of variation less than 0.55%for both intra-and intergroups.This method has the potential to support early diagnosis,epidemiological monitoring and effective prevention and control of the disease.
Keywords：Goose reovirusGRV and NGRVdye-based methodRT-qPCRMelting curve
Publication Date：2025-06-18
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：9( 96-104 )