| 林秀银 1
，温肖会 2，吕殿红 2，翟少伦 2，霍 玮 2，翟 颀 2，魏文康 2，杨彩娟 3.广东地区猪群新发塞内卡病毒流行株的分离鉴定及遗传进化分析[J].广东农业科学,2019,46(6):125-132 |
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广东地区猪群新发塞内卡病毒流行株的分离鉴定及遗传进化分析 |
| Isolation, Identification and Genetic Evolution Analysis of New Epidemic Strains of Senecavirus A in Guangdong Province |
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| DOI：10.16768/j.issn.1004-874X.2019.06.017 |
| 中文关键词: 塞内卡病毒 口蹄疫 猪原发性水疱病 分离鉴定 序列分析 |
| 英文关键词: Senecavirus A foot-and-mouth disease virus porcine idiopathic vesicular disease isolation and identification sequence analysis |
| 基金项目:广东省农业科学院学科团队建设（201634TD）；广东省农业农村厅重大动物疫病防控技术研究转化和推广（粤农计〔2018〕54 号）；广东省科技计划项目（2016B020234006） |
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| 中文摘要: |
| 【目的】检测广东两猪场新发的鼻镜与蹄冠水疱病病原，获得新发塞内卡病毒（Senecavirus A，
SVA）流行株，为后续研究提供材料。【方法】应用 SVA RT-PCR、荧光 PCR 等方法对采集的病料进行初步筛
查，再将水泡皮进行处理接种于 ST 细胞、BHK-21 细胞上，用 SVA 特异性引物对细胞上清液进行扩增，测序
并分析序列，采用蔗糖浓度梯度方法纯化病毒进行电子显微镜形态观察，并对病毒分离株的 TCID50 进行测定。
【结果】 经 SVA RT-PCR 与荧光 PCR 方法初步诊断该猪群为 SVA 感染；细胞接毒培养发现，自第 6 代起，
细胞出现典型 CPE，电镜观察可见 25 nm 左右的病毒颗粒，扩增序列提交 NCBI Blastn 比对鉴定为 SVA；测得
毒株 SVA CH-GDBL1-2016 的 TCID50 为 106.8，毒株 SVA CH-GDBL2-2016 的 TCID50 为 106.7。【结论】SVA
CH-GDBL1-2016 和 SVA CH-GDBL2-2016 可为后续塞内卡病毒病的诊断和疫苗研制提供材料基础。 |
| 英文摘要: |
| 【Objective】 The pathogen of blistering disease with nasal and coronet in two swine farms in Guangdong
province was detected, and new strains of Senecavirus A (SVA) were obtained, which provided materials for subsequent
research. 【Method】 RT-PCR and real-time RT-PCR were used to screen the clinical samples. The clinical samples
were isolated, treated and cultured with ST cells and BHK-21 cells. The virus was purified by sucrose density gradient centrifugation method and observed by electron microscopy. The specific primers of SVA were designed to amplify the cell
supernatant, and sequenced for sequence analysis. The TCID50 of the virus isolated was determined. 【Results】 The swinery
were diagnosed as SVA by RT-PCR and real-time RT-PCR. It was found that the cultured cells had CPE from the 6th
generation. The virion observed by electron microscopy was about 25 nm. The amplified sequences were submitted to the NCBI
Blastn, which were identified as SVA. The TCID50 result of the strain SVA CH-GDBL1-2016 was 106.8 and the TCID50 result of
the strain SVA CH-GDBL2-2016 was 106.7.【Conclusion】 The strains SVA CH-GDBL1-2016 and SVA CH-GDBL2-2016
provide a material basis for the subsequent diagnosis and vaccine development of Senecavirus disease. |
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