鸭瘟病毒强弱毒株PCR检测方法的建立

发布时间：2018-01-19 00:35

本文关键词： 鸭瘟病毒强毒弱毒聚合酶链反应　出处：《动物医学进展》2017年08期 　论文类型：期刊论文

【摘要】：根据基因库中鸭瘟病毒强毒株和弱毒株UL2基因保守区设计特异性引物,优化PCR反应的引物浓度和退火温度等,初步建立了可同时鉴别鸭瘟病毒强毒株和弱毒株的PCR检测方法,并对建立的方法进行了敏感性、特异性验证和临床样品检测。该PCR检测方法最低能检出1pg的鸭瘟病毒强毒和弱毒DNA模板。对鸭瘟病毒强毒和弱毒模板的检测,得到了与试验设计相符的827bp(强毒)和299bp(弱毒)的扩增条带,而对鸭副黏病毒、鸭坦布苏病毒、鸭圆环病毒、番鸭细小病毒、鸭Ⅰ型肝炎病毒、禽流感病毒和小鹅瘟病毒等病原体的检测均为阴性。说明建立了一种敏感性高、特异性好的鉴别鸭瘟病毒强毒和弱毒的PCR检测方法。
[Abstract]:Specific primers were designed according to the conserved region of UL2 gene of highly and attenuated strains of duck plague virus in the gene bank to optimize the concentration of primers and annealing temperature of PCR reaction. A PCR method for the identification of both virulent and attenuated strains of duck plague virus was established, and the sensitivity of the established method was evaluated. The PCR method can detect 1 PG of highly toxic and attenuated DNA templates of duck plague virus, and detect the highly virulent and attenuated templates of duck plague virus. The amplified bands of 827bp (strong virus) and 299bp( attenuated virus) were obtained in accordance with the experimental design. The results showed that the amplified bands were suitable for duck paramyxovirus, duck tamosu virus, duck circovirus and muscovy duck parvovirus (Muscovy duck parvovirus). The detection of duck hepatitis I virus, avian influenza virus and goose plague virus were all negative, which indicated that a highly sensitive and specific PCR method for the identification of duck plague virus was established.
【作者单位】：广西壮族自治区兽医研究所广西兽医生物技术重点实验室;广西玉林市动物疫病预防控制中心;
【基金】：广西科技项目(桂科攻10100014-5) 国家”万人计划”领军人才专项(2016-37) 广西特聘专家专项(2011B020)
【分类号】：S852.65
【正文快照】： 鸭瘟是由鸭瘟病毒(Duck plague virus,DPV)引起鸭的一种急性败血性传染病,发病率和病死率都甚高,是危害养鸭业最为严重的传染病之一[1]。近年来,随着商品肉鸭集约化饲养范围的不断扩大,该病在商品鸭群中的发病呈明显上升趋势,未免疫鸭群的病死率接近100%,而且在同一地区或同一

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