鸽圆环病毒TaqMan MGB探针荧光定量PCR检测方法的建立及全基因组分析

鸽业是我国传统的养殖业之一，具有悠久的历史和广泛的区域分布。然而，随着时代的发展，养殖业模式逐渐向集约化方式转变，在密集环境中，鸽群易受到传染性病原体的侵袭，造成大范围的感染和传播。其中，鸽圆环病毒（Pigeon circovirus，PiCV，）是一种对鸽具有高传染性并能引起免疫抑制的病原体，可导致鸽羽毛异常、厌食、腹泻、生长迟缓和继发感染（如免疫器官衰退引发的沙门氏菌等多种条件致病病原体感染），对养鸽业造成重大的经济损失^。。目前，尚无体外细胞分离培养PiCV的有效方法。核酸检测是实现快速诊断和监测PiCV的重要手段。本研究拟通过比对不同PiCV 的基因序列获取其保守区域，设计特异性检测引物和MGB 公司探针，优化反应条件，建立一种高敏感性、强特异性、良好重复性和临床适用性的荧光定量聚合酶链反应方法。为PiCV 的早期诊断、科学防控及流行病学调查提供一种基于MGB 公司探针建立的新型检测技术。

Pigeon farming is one of the traditional breeding industries in China, boasting a long history and wide regional distribution. However, with the advancement of the times, the breeding mode has gradually shifted towards intensive farming. In dense breeding environments, pigeon flocks are highly susceptible to invasion by infectious pathogens, resulting in large-scale infection and transmission. Among these pathogens, Pigeon circovirus (PiCV) is a highly infectious agent that causes immunosuppression in pigeons. It can induce abnormal feathers, anorexia, diarrhea, growth retardation, and secondary infections, including infections by multiple opportunistic pathogens such as Salmonella triggered by immune organ atrophy, leading to substantial economic losses for the pigeon farming industry. Currently, there are no effective methods for in vitro isolation and culture of PiCV. Nucleic acid detection serves as an important tool for rapid diagnosis and monitoring of PiCV. This study intends to obtain the conserved regions of PiCV by aligning the gene sequences of different PiCV strains, design specific detection primers and MGB company probes, optimize reaction conditions, and establish a quantitative fluorescence polymerase chain reaction (qPCR) assay with high sensitivity, strong specificity, good reproducibility and clinical applicability. This will provide a novel detection technology based on MGB company probes for the early diagnosis, scientific prevention and control, and epidemiological investigation of PiCV.