检测新型鹅细小病毒的半套式PCR方法的建立及应用  

Development and application of a semi-nested PCR assay for detecting novel goose parvovirus

作　　者：高沙沙 李鹏飞 孙大鹏 张瑞华[1] 陈君豪[1] 蓝晶晶 谢之景[1] 姜世金[1] GAO Sha-sha;LI Peng-fei;SUN Da-peng;ZHANG Rui-hua;CHEN Jun-hao;LAN Jing-jing;XIE Zhi-jing;JIANG Shi-jin(College of Veterinary Medicine of Shandong Agricultural University,Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention,Shandong Provincial Engineering Technology Research Center of Animal Disease Control and Prevention Taian 271018,China)

机构地区：[1]山东农业大学动物医学院/山东省动物生物工程与疾病防治重点实验室/山东省畜禽疫病防控工程技术研究中心,山东泰安271018

出　　处：《中国预防兽医学报》2018年第8期702-705,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家重点研发计划项目(2016YFD05008);山东省双一流建设项目(SYL2017YSTD11);山东省现代农业产业技术体系家禽创新团队(SDAIT-11-15)

摘　　要：2015年以来山东、江苏等地的樱桃谷肉鸭暴发了一种由新型鹅细小病毒(NGPV)引起的短喙-侏儒综合征(BADS)的疾病,造成了严重的经济损失。为探究该病毒在鸭体内分布规律,本研究根据Gen Bank已登录的NGPV的VP3基因设计了3条引物,建立了检测NGPV的半套式PCR方法。该方法对其它常见鸭病原微生物无特异扩增,特异性好;最低可检出100拷贝/μL的病毒核酸,灵敏度高。采用建立的半套式PCR方法对山东、江苏两地8个樱桃谷肉鸭鸭群120只病死鸭的心脏、肝脏、脾脏、肺脏、肾脏、胸腺、法氏囊、胰腺、脑和胆汁进行检测,结果显示患病鸭群的群体检出率为100%,个体检出率为80.8%(97/120);10种组织脏器中均检出NGPV,其中心脏检出率最高为93.8%,脑组织检出率最低为9.3%。本研究首次对NGPV在鸭体内10个脏器组织的分布进行统计分析,证明了NGPV对樱桃谷鸭具有广泛的组织嗜性。A novel goose parvovirus-related virus（NGPV） causing the disease of duck beak atrophy and dwarfism syndrome（BADS） in Cherry Valley ducks has broken out in Shandong and Jiangsu provinces since 2015, which caused great economic losses to waterfowl pouftry industry. To explore the distribution of NGPV in different organs of infected ducks, a semi-nested PCR assay was developed with 3 primers designed based on the NGPV sequences available in GenBank and applied in detection of clinical duck samples. The PCR assay was high specificity and sensitivity with the minimum detection limit of 100 copies for the viral DNA. Using the semi-nested PCR assay, DGPV was detected in all the 8 Cherry Valley meat duck flocks, and 97 ducks（80.8%） were DGPV positive. In the NGPV-positive ducks, the virus was detected in all the ten tested tissue samples（heart, liver,spleen, lung, kidney, pancreas, bile, thymus, brain, and bursa of Fabricius）, with the highest detection rate in heart（93.8%） and the lowest detection rate in brain（9.3%）. In this study, we first detected and analyzed ten different organ tissues of dead ducks in Shandong and Jiangsu province, which demonstrated that NGPV has an extensive tropism to tissues in Cherry Valley ducks.

关 键 词：新型鹅细小病毒 半套式PCR 检出率 

分 类 号：S852.65[农业科学—基础兽医学]