Two pairs of primers were designed to bind conserved genomic regions of goose parvovirus (GPV) and goose astrovirus (GAstV) to establish a simple, sensitive, and highly specific duplex quantitative PCR (qPCR) method to simultaneously detect the two viruses. The duplex qPCR can distinguish GPV (melting point: 82.1 °C) and GAstV (melting point: 79.8 °C) by the peaks of their individual melting curves. Mixed testing with other waterfowl viruses produced no nonspecific peaks. The established standard curves showed good linear relationships (R² > 0.997) and the limits of detection (LOD) for GPV and GAstV were 5.74 × 10¹ and 6.58 × 10¹ copies/μL, respectively. Both intra- and inter-assay coefficients of variation were <2%, indicating that the method has good repeatability. Twenty tissue samples from diseased geese were examined with the duplex qPCR assay and conventional PCR. Duplex qPCR showed positive rates of 25% for GPV and 45% for GAstV, and the positive rate for GPV and GAstV coinfection was 15%, slightly higher than the results for conventional PCR. These results indicated that this duplex qPCR method is highly sensitive, specific, and reproducible, and is suitable for epidemiological studies to effectively control the transmission of GPV and GAstV.

设计了两对引物以结合鹅细小病毒（GPV）和鹅星状病毒（GAstV）的保守基因组区域，以建立一种简单，灵敏且高度特异性的双重定量PCR（qPCR）方法来同时检测这两种病毒。双工qPCR可以通过各自的熔解曲线的峰来区分GPV（熔点：82.1°C）和GAstV（熔点：79.8°C）。与其他水禽病毒的混合测试没有产生非特异性峰。建立的标准曲线显示出良好的线性关系（R ²  > 0.997），并且GPV和GAstV的检出限（LOD）为5.74×10 ¹和6.58×10 ¹拷贝数/μL。测定内和测定间变异系数均<2％，表明该方法具有良好的重复性。用双重qPCR测定法和常规PCR检查了二十只来自患病鹅的组织样品。双重qPCR显示GPV的阳性率为25％，GAstV的阳性率为45％，而GPV和GAstV合并感染的阳性率为15％，略高于常规PCR的结果。这些结果表明，这种双工qPCR方法具有高度的敏感性，特异性和可重复性，适用于流行病学研究以有效控制GPV和GAstV的传播。