小鹅瘟病毒荧光RPA恒温快速检测方法的建立与应用  

Rapid Detection of Goose Parvovirus Based on Isothermal Recombinase Polymerase Amplification Assay

作　　者：高远 徐龙涛 冯宗玲 曲光刚[4] 李本科 GAO Yuan;XU Long-tao;FENG Zong-ling;QU Guang-gang;LI Ben-ke(Animal Husbandry Development Center of Rongcheng City, Rongcheng, Shandong 264399, China;Qilu Animal Health Products Co., LTD., Jinan 250100, China;Binzhou Institute of Science and Technology Innovation and Development, Binzhou, Shandong 256600, China;Shandong Binzhou Animal Science and Veterinary Medicine Academy, Binzhou, Shandong 256600,China;Binzhou Agricultural Technology Extension Center, Binzhou, Shandong 256600, China)

机构地区：[1]荣成市畜牧业发展中心,山东荣成264399 [2]齐鲁动物保健品有限公司,济南250100 [3]滨州市科技创新发展研究院,山东滨州256600 [4]山东省滨州畜牧兽医研究院,山东滨州256600 [5]滨州市农业技术推广中心,山东滨州256600

出　　处：《中国兽药杂志》2022年第3期1-10,共10页Chinese Journal of Veterinary Drug

基　　金：山东省重点研发计划(重大科技创新工程)项目(2019 JZZY010720)。

摘　　要：为快速检测小鹅瘟病毒,以小鹅瘟病毒VP3基因保守片段为靶点,利用重组酶聚合酶扩增技术(RPA)建立了一种准确高效的小鹅瘟病毒RPA恒温快速检测方法,对检测方法的灵敏度、特异性进行评价,并与传统PCR和Real-time PCR方法进行比较。结果表明,该检测方法可检测到10 copies/μL的病毒核酸,具有较高的灵敏度;只特异性地扩增小鹅瘟病毒,而与鹅副黏病毒、鹅源鸭瘟病毒、小鹅流行性感冒病毒、鹅副伤寒病毒、大肠杆菌和曲霉菌均未发生交叉反应,特异性良好;变异系数低于6%,具有很好的重复性;临床样品检测中与Real-time PCR符合率为99%。该方法可以很好地应用于小鹅瘟的大规模临床样本检测,为小鹅瘟病毒的高通量检测和流行病学调查提供技术手段。Based on the recombinase polymerase amplification(RPA)technology,and the conserved fragment of VP3 gene as the target,an accurate and efficient constant temperature method for detecting goose parvovirus was established.The sensitivity and specificity of the detection method were evaluated and compared with traditional PCR and Real-time PCR.Results showed that the method was highly sensitive with a detection limit of 10 copies/μL of viral nucleic acid.This method also exhibited a good specificity without any cross reaction with goose paramyxovirus,goose distemper virus,gosling influenza virus,goose paratyphoid virus,E.coli and the coefficient of variation of this method was less than 6%,which indicated good repeatability.The test of clinical samples showed that the coincidence rate between this method and real-time PCR was 99%.This method can be applied to the detection of gosling disease in large scale clinical samples,and provide a novel method for the high throughput detection and epidemiological investigation of gosling disease virus.

关 键 词：小鹅瘟病毒 重组酶聚合酶扩增技术(RPA) 恒温快速检测 

分 类 号：S852.65[农业科学—基础兽医学]