摘要
近年来,我国大陆地区暴发了一种由新型鹅细小病毒（NGPV）感染引起的鸭“短喙侏儒综合征”的疫病。为了建立一种特异性好、灵敏度高适合临床大批量样本检测的方法,本试验将鸭源NGPV的VP3保守区域1段长390bp的序列克隆制成核酸探针。特异性试验结果表明,制备的该核酸探针特异性高,只与鹅细小病毒（GPV）反应;灵敏性试验结果表明,针对NGPV最低检出量约为6pg。应用该核酸探针对来自山东、江苏、安徽3省的87只疑患“短喙长舌-侏儒综合征”的樱桃谷肉鸭的870份组织脏器进行检测,结果个体阳性率为89.7%（78/87）,检测的10种组织脏器中均有NGPV检出,说明该病毒对樱桃谷鸭具有广泛的组织嗜性。同时,利用PCR方法对采集的样品进行检测,比较发现与核酸探针法的阳性个体检出率及阳性个体检出符合率为100%。以上结果表明,制备的NGPV核酸探针特异性强,灵敏性高,适合不同来源的NGPV临床批量诊断和流行病学调查。
In resent years,a duck infectious disease characterized by ＂short beak and dwarfism syn- drome （SDBS）＂, caused by a novel goose parvovirus（NGPV）, broke out in Pekin ducks in China's Mainland. In order to establish a method with high specificity and sensitivity for high-volume sample detection,a length of 390bp DNA that located in VP3 of duck-origin NGPV was cloned to make DNA probe. The DNA probe was demonstrated with high specificity and the probe can only com- bine and react to goose parvovirus（GPV）rather than other pathogen DNAs. The specificity test showed that the minimum detection limit to be 6pg viral DNA of NGPV. Using the DNA probe,a total of 870 tissues and organs of 87 cherry valley ducks with typical clinical symptoms of SBDS from Shandong,Jiangsu and Anhui provinces were tested. The results indicated that the individual positive ratio of NGPV was 89.7%（78/87）,and NGPV was detected in all the 10 kinds of tissue and organs,indicating that NGPV has a wide tissue tropism in cherry valley ducks. At the same time,all the collected samples were detected by PCR method. By comparison of the DNA probe and the PCR,we found that the positive individual detection rate and positive individual detection coincidence rate were 100%. The results showed that the prepared NGPV DNA probe had high specificity and sensitivity and was suitable for clinical diagnosis and epidemiological investigation of NGPV from different sources.
作者
李鹏飞
张瑞华
宋莎莎
王玮
陈君豪
蓝晶晶
谢之景
姜世金
LI Peng-fei,ZHANG Rui-hua, SONG Sha-sha, WANG Wei, CHEN Jun-hao, LAN Jing-jing, XIE Zhi- jing,JIANG Shi-jin(Shandong Provincial Engineering Technology Research Center of Animal Disease Control and Prevention/Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention ,College of Veterinary Medicine, Shandong Agricultural Uni- versity, Taian, Shandong 2 71018, China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2018年第11期2073-2077,共5页
Chinese Journal of Veterinary Science
基金
国家重点研发计划资助项目(2016YFD0500805-5)
山东省双一流建设资助项目
山东省农业重大应用技术创新课题资助项目
山东省现代农业产业技术体系家禽创新团队资助项目(SDAIT-11-15)
