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.

Key words: Goose reovirus, GRV and NGRV, dye?based method, RT?qPCR, Melting curve

摘要： 鹅呼肠孤病毒（Goose reovirus, GRV）包括经典型（GRV）和新型鹅呼肠孤病毒（Novel Goose reovirus, NGRV），两者在流行病学特征和病毒基因序列上存在差异。目前针对这两种类 型病毒的敏感、特异的检测方法尚未完善，使得准确诊断和及时干预变得十分困难。为了区分 GRV和NGRV，建立了一种基于染料法（SYBR Green I）的双重RT?qPCR检测方法。结果显示： 该方法通过熔解曲线可区分GRV（79.5℃）和NGRV（86.5℃），对常见的水禽流行疫病均未出现 扩增，最低检出为6.466×101拷贝/μLGRV和4.979×101拷贝/μLNGRV，组内与组间变异系数均 小于0.55%。结果表明，这个研究所建立基于染料法的双重RT?qPCR检测方法可以快速准确 地确定感染鹅群中不同型的GRV，为疾病的早期诊断、流行病学监测和有效防控提供支持。

关键词: 鹅呼肠孤病毒, GRV和NGRV, 染料法, RT?qRCR, 熔解曲线