【摘要】 圆圈病毒禽源1型(GyVg1)和圆圈病毒人源1型(GyH1)是两种新发现的可引起鸡腺胃炎相关症状的环形病毒，目前在全球多个地区均有检测报道。本研究旨在建立一种能快速同时鉴别GyVg1和GyH1的二重荧光定量PCR方法。使用GenBank中现有的GyVg1和GyH1基因组序列，基于其保守区分别设计相应的特异性引物和探针，经过优化反应条件，建立了GyVg1和GyH1的二重荧光定量PCR检测方法。应用该检测方法对2023年在广西南宁8个地区采集的256份临床样本进行检测，与常规PCR检测和测序结果进行分析比较，进一步验证该方法的有效性。结果显示，所建立的二重荧光定量PCR方法可在1 h内鉴定出GyVg1和GyH1,两套引物探针均可有效扩增，且与其他病原体无交叉反应；检测下限为7.5拷贝/μL,标准曲线相关系数>0.99,批内和批间变异系数<0.45%。临床样品检测结果显示，当模板量≥100.0拷贝/μL时，二重荧光定量PCR测定与常规PCR测定之间的一致性分别为94.3%(GyVg1)和100%(GyH1)。综上，所建立的二重荧光定量PCR方法特异性强、灵敏度高、重复性好，适用于GyVg1和GyH1的快速鉴别检测。更多还原

【Abstract】 Gyrovirus galgal(GyVg1) and gyrovirus homsal(GyH1) are two newly discovered circoviruses that can cause symptoms related to transmissible viral proventriculitis of chickens.These viruses have been reported in various regions worldwide.This research aims to establish a duplex real-time PCR assay capable of identifying and detecting GyVg1 and GyH1.Specific primers and probes were designed based on the conserved regions of GyVg1 and GyHl respectively using all genome sequence data currently available in GenBank.After optimizing reaction conditions,the duplex real-time PCR detection method was established and further validated by comparing it with a conventional PCR assay and sequencing results from an analysis of 256 clinical samples collected in2023 across eight regions of Nanning,Guangxi.The results showed that GyVg1 and GyH1 could be identified in 1 h by the duplex real-time PCR assay and two pairs of primer probes can amplify effectively but there is no any cross reaction with other pathogens.Besides,the detection limit was determined to be 7.5 copies/μL.The correlation coefficient of standard curves exceeded 0.99,and CV for intra-and inter-assay was less than 0.45%.Based on clinical performance,when the quantity of template was greater than or equal to 100 copies,the agreements between the duplex realtime PCR assay and the conventional PCR assay were 94.3%(GyVg1) and 100%(GyH1),In conelusion,the newly developed duplex real-time PCR assays exhibited good specificity,sensitivity and repeatability,which could contribute to the rapid detection and differentiation of GyVg1 and GyH1.更多还原