摘要
2015年暴发了一种由新型鹅细小病毒(NGPV)引起的鸭喙萎缩-侏儒症(BADS)的疾病,造成了严重的经济损失。本研究根据GenBank中NGPV相对保守的VP3基因序列,设计1对特异性引物和1条TaqMan探针,并对其反应条件进行优化,建立了一种检测NGPV的TaqMan荧光定量PCR方法。敏感性试验结果表明,该方法检测敏感性达到37.2拷贝/μL,比常规PCR灵敏度高100倍,而且该方法对鸭其他主要病原核酸检测均为阴性,具有良好的特异性。批内和批间的检测变异系数均小于等于2.2%。对收集的50份病料进行检测,本研究建立的荧光定量PCR方法NGPV检出率为78%,普通PCR方法为50%,荧光定量PCR方法的敏感性明显高于普通常规PCR方法。研究结果表明该方法具有操作简便、敏感性高和特异性强的特点,可用于NGPV的快速诊断。
A newly emerged duck beak atrophy and dwarfism syndrome(BADS)caused by a novel goose parvovirus-related virus(NGPV)outbroke in 2015,leading to a great economic loss for waterfowl poultry industry.For detection of NGPV infection,a TaqMan-based real-time PCR assay was developed using a pair of primers and a TaqMan probe that were designed and synthesized according to the conserved VP3 gene sequences.The real-time PCR assay specifically detected NGPV and had no cross-reaction with other duck viruses.The detection limit of this assay was 37.2 viral copies,which was 100 times higher than that of the conventional PCR,indicating its significantly higher sensitivity than that of conventional PCR method.The coefficients of variability for intra-assay and inter-assay were both less than 2.2%.Furthermore,the real-time PCR positive rate of the tested NGPV samples was 78%,while that of the conventional PCR was 50%.The results from this study indicated that the real-time PCR method developed here was simple,sensitive and specific and could be used for rapid laboratory diagnosis and monitoring of NGPV infection.
作者
曹旭阳
王宇飞
张志飞
魏笑笑
秦立廷
陈佶慧
滕巧泱
杨健美
刘芹防
陈鸿军
李泽君
李景环
李雪松
CAO Xu-yang;WANG Yu-fei;ZHANG Zhi-fei;WEI Xiao-xiao;QIN Li-ting;CHEN Ji-hui;TENG Qiao-yang;YANG Jian-mei;LIU Qin-fang;CHEN Hongjun;LI Zejun;LI Jing-huan;LI Xue-song(College of Life Science and Technology,Inner Mongolia Normal University,Hohhot 010022,China;Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China;Shandong New Hope Liuhe Group Co.,Ltd,Qingdao 266000,China)
出处
《中国动物传染病学报》
CAS
北大核心
2020年第5期31-36,共6页
Chinese Journal of Animal Infectious Diseases
基金
国家重点研发计划“家禽重要疫病诊断与检测新技术研究”(2016YFD0500800)
国家自然科学基金项目(31702237,31771000)
上海市自然科学基金项目(17ZR1437400)
上海科委创新行动计划农业项目(17391901700)。
关键词
新型鹅细小病毒
荧光定量PCR
检测
Novel goose parvovirus-related virus
real-time PCR
detection
