Diagnosis of Duckling Short Beak and Dwarfism Syndrome by Indirect Immunofluorescence Assay
-
摘要: 应用抗鹅细小病毒（GPV）单克隆抗体（Mab E16）为一抗，建立检测鸭短嘴矮小综合征病毒（SBDS-GPV）的间接免疫荧光方法（IFA），并对人工感染SBDS-GPV的发病鸭和临床送检鸭短嘴矮小综合征（SBDS）病鸭的肝脏、脾脏、肾脏和胰腺进行检测。结果显示该方法具有良好的特异性，能识别鹅细小病毒抗原呈现亮绿色特异荧光，而与MPV、MDRV、NDRV、DHV、DPV、PMV等其他鸭病病原均无交叉反应；应用该方法检测SBDS-GPV人工感染发病鸭和SBDS临床病鸭组织，均不同程度出现亮绿色荧光，其中人工感染发病鸭阳性率100%（5/5），临床送检病鸭阳性率91.7%（11/12）。表明建立的IFA具有快速、特异等优点，可用于SBDS的快速诊断。Abstract: An indirect immunofluorescence assay (IFA) was established using the monoclonal antibodies (Mab E16) against goose parvovirus (GPV) for identification of the short beak and dwarfism syndrome (SBDS)-GPV in ducks. Liver, spleen, kidney and pancreas of the ducklings challenged with SBDS-GPV and the natural infection cases were tested by the new IFA methodology. The results showed that the newly developed IFA was specific, giving positive reaction to GPV only and negative to other viruses of MPV, MDRV, NDRV, DHV, DPV and PMV. Five out of 5 challenged cases and 11 out of 12 natural infection cases showed a positive reaction with bright green fluorescence glow on IFA. Consequently, the new method could be applicable for detecting and diagnosing SBDS in ducks with high specificity, accuracy and rapidity.
-
图 1 IFA检测结果
注：A为阴性，B为肝脏，C为脾脏，D为肾脏，E为胰腺。
Figure 1. Test results using newly developed IFA methodology
表 1 单抗E16稀释度和IFA特异性测定结果
Table 1. Mab E16 concentration and specificity of IFA
单抗稀释度 SBDS-GPV MD-GPV MPV MDRV NDRV DHV 正常鸭胚 1:50 +++ +++ - - - - - 1:100 +++ +++ 1:200 ++ ++ 注：“-”表示50个视野均未见荧光灶；“++”表示1个视野见1~2个荧光灶；“+++”表示1个视野见5个以上荧光灶。 
下载: 导出CSV
表 2 SBDS病鸭的IFA检测结果
Table 2. IFA results on ducks suspected with SBDS
组织 不同日龄半番鸭/d 不同日龄樱桃谷鸭/d 15 7 12 6 13 6 35 16 22 27 17 18 肝脏 ++ ++ ++ + ++ + - ++ + + + + 脾脏 + + ++ + + + - + - - - - 胰腺 +++ ++ +++ + ++ + - +++ + ++ + ++ 肾脏 +++ ++ ++ + ++ + - ++ + ++ + ++ 判定结果 +++ ++ +++ + ++ + - +++ + ++ + ++ 注：-为50个视野未见荧光灶，+为10视野见1~2个荧光灶，++为1个视野见1~2个荧光灶，+++为1个视野见5个以上荧光灶。
下载: 导出CSV
-
[1] 陈少莺, 程晓霞, 陈仕龙, 等.半番鸭新型小鹅瘟病毒病研究简报[J].福建农业科技, 2015, 46(7):23-25. https://www.wenkuxiazai.com/doc/994fd43cba0d4a7303763a6e.html [2] 吴南洋, 黄梅清, 陈少莺, 等.鸭短嘴矮小综合征的流行病学调查[J].中国家禽, 2016, 38(4):66-67. http://www.doc88.com/p-4117669382351.html [3] CHEN S L, WANG S, CHENG X X, et al. Isolation and characterization of a distinct duck-origin goose parvovirus causing an outbreak of duckling short beak and dwarfism syndrome in China[J]Archives of Virology, 2016, 161(9):1-10. doi: 10.1007/s00705-016-2926-4.pdf [4] WANG S, CHENG X X, CHEN S L, et al. Identification of a novel goose parvovirus (GPV) recombinant associated with short beak and dwarfism syndrome in Mainland China, 2015[J]. Infection Genetics & Evolution, 2016, 41:289. https://www.deepdyve.com/lp/elsevier/identification-of-a-novel-goose-parvovirus-gpv-recombinant-associated-8AfmWX0U0N [5] WANG S, CHENG X X, YU B, et al. A simple, polymerase chain reaction and restriction fragment length polymorphism-aided diagnosis method for short beak and dwarfism syndrome in ducklings[J]. Infect Genet Evol, 2017, 53:85-88. doi: 10.1016/j.meegid.2017.05.014 [6] XIAO S, CHEN S, CHENG X, et al. The newly emerging duck-origin goose parvovirus in China exhibits a wide range of pathogenicity to main domesticated waterfowl[J]. Veterinary Microbiology, 2017, 203:252. doi: 10.1016/j.vetmic.2017.03.012 [7] 殷震, 刘景华.动物病毒学:第二版[M].北京:科学出版社, 1997:223-224. [8] 方定一. "小鹅瘟"的介绍[J]. 中国畜牧兽医, 1962, (8): 19-20. [9] 程晓霞, 陈少莺, 朱小丽, 等.番鸭小鹅瘟病毒的分离与鉴定[J], 福建农业学报, 2008, 23(4):355-358. http://www.fjnyxb.cn/CN/abstract/abstract281.shtml [10] 朱小丽, 陈少莺, 程晓霞, 等.小鹅瘟病毒单克隆抗体的制备及特性鉴定[J].中国动物传染病学报, 2011, 19(6):20-24. http://kns.cnki.net/KCMS/detail/detail.aspx?filename=zsjb201106006&dbname=CJFD&dbcode=CJFQ -
点击查看大图
图(1) / 表(2)
计量
- 文章访问数: 1304
- HTML全文浏览量: 156
- PDF下载量: 89
- 被引次数: 0
