摘 要
猪流行性腹泻（PED）是猪流行性腹泻病（PEDV）所造成的一种高程度的传染性肠炎。该病由最初的流行季（11月份到翌年3月）发展为一种可在每年一度流行的疾病，在猪体表面的阳性率和肉品检出率分别为52.24%和1.87%。吉林地区现有的 PED疫情尚不能得到有效的预防，因此，本区域 PEDV感染与我国的 PED病毒感染存在一定的差别。为了进一步了解吉林农村 PEDV的基因变异和分布特点，对吉林农村 PEDV的预防和控制具有较好的指导意义，同时还应及时采取相应的应急处理方法，以最大限度的降低生产成本和降低生产经营的危险。
本文采用 Primer5.0的方法，利用 GenBank公司发布的 PEDV基因序列，对2016年2-2018年2月份在吉林市和舒兰，蛟河，永吉，桦甸，盘石等6个县46家养猪310个品种进行RT-PCR分析， PCR结果均为阳性，并将 PCR结果进行分析，并将 PCR结果与上海生工区进行比较。经检验，46家养猪中19家为 PEDV感染的养猪，其检出率达41.3%;310家养猪中，检出 PEDV的80家，其检出率为25.8%；永吉养猪的阳性率分别为58.33%和16.67%，桦甸地区为16.67%；在各季中，各地区的感染率以冬季最高。结果表明： PED发病率与各地区、各猪场免疫程序、季节和温度变化有密切的联系，且与我国大部分的病原有密切的联系。
关键词：猪流行性腹泻病毒 分子生物学 流行病学
Abstract
Porcine epidemic diarrhea (PED) is a highly infectious enteritis caused by porcine epidemic diarrhea disease (PEDV). The disease developed from the initial epidemic season (November to March of the next year) to a disease that can be epidemic once a year. The positive rate on the surface of pigs and the detection rate of meat products were 52.24% and 1.87%, respectively. The current epidemic situation of PED in Jilin cannot be effectively prevented. Therefore, there is a certain difference between PEDV infection in this region and that in China. In order to further understand the genetic variation and distribution characteristics of rural PEDV in Jilin Province, it has a good guiding significance for the prevention and control of rural PEDV in Jilin Province. At the same time, corresponding emergency treatment methods should be taken in time to minimize production costs and reduce the risk of production and operation.
In this paper, Primer 5.0 method was used to analyze 310 breeds of 46 pigs raised in Jilin City and Shulan, Jiaohe, Yongji, Huadian, Panshi and other six counties from February 2016 to February 2018 by using the PEDV gene sequence released by GenBank. The PCR results were all positive, and the PCR results were analyzed and compared with those in Shanghai production area. After inspection, 19 out of 46 pigs were infected with PEDV, and the detection rate was 41.3%; Among 310 pigs, 80 were found to have PEDV, with a detection rate of 25.8%; The positive rates of Yongji pigs were 58.33% and 16.67%, respectively, and 16.67% in Huadian area; In each season, the infection rate in each region was the highest in winter. The results showed that the incidence rate of PED was closely related to the immunization program, season and temperature changes in various regions and pig farms, and was closely related to most pathogens in China.
Key words: Porcine epidemic diarrhea virus Molecular Biology epidemiology
目 录
摘 要 I
Abstract II
1 猪流行性腹泻的研究进展 1
1.1 病原学 1
1.1.1 病原分类 1
1.1.2病原形态 1
1.1.3病毒基因组 1
1.1.4病毒理化特征 1
1.1.5病毒抗原性 1
1.1.6 病毒培养特性 2
1.2 流行病学 2
1.2.1 易感动物 2
1.2.2传播途径 2
1.2.3传染源 2
1.3流行动态 2
1.3.1 国外流行情况 3
1.4发病机理 4
1.5临床症状 4
1.5.1 哺乳仔猪 4
1.5.2断奶仔猪及保育仔猪 5
1.5.3生长育肥猪 5
1.5.4种母猪 5
1.5.5种公猪 5
1.6病理变化 5
1.6.1 眼观病理变化 5
1.6.2 镜观病理变化 5
1.7诊断方法 6
1.7.1 病原学诊断 7
1.7.2 血清学诊断 7
1.8 PED的防治 8
1.8.1 疫苗研究与应用 8
1.8.2 PED 的防控 10
2 吉林市猪流行性腹泻分子流行病学调查 10
2.1 试验材料 11
2.1.1 主要试剂 11
2.1.2 主要仪器 11
2.1.3 引物 12
2.1.4毒株 12
2.2 试验方法 12
2.2.1病料采集 12
2.2.2 RNA 提取 12
2.2.3 电泳 13
2.2.4凝胶回收 13
2.2.5克隆与测序 13
3 结果 14
3.1 PCR鉴定结果 14
3.2 吉林省规模化猪场不同季节仔猪病料 PCR 结果 15
3.3 测序结果 15
4 讨论 16
5 结论 17
参考文献 18
致 谢 20
猪流行性腹泻（PED）是猪流行性腹泻病（PEDV）所造成的一种高程度的传染性肠炎。该病由最初的流行季（11月份到翌年3月）发展为一种可在每年一度流行的疾病，在猪体表面的阳性率和肉品检出率分别为52.24%和1.87%。吉林地区现有的 PED疫情尚不能得到有效的预防，因此，本区域 PEDV感染与我国的 PED病毒感染存在一定的差别。为了进一步了解吉林农村 PEDV的基因变异和分布特点，对吉林农村 PEDV的预防和控制具有较好的指导意义，同时还应及时采取相应的应急处理方法，以最大限度的降低生产成本和降低生产经营的危险。
本文采用 Primer5.0的方法，利用 GenBank公司发布的 PEDV基因序列，对2016年2-2018年2月份在吉林市和舒兰，蛟河，永吉，桦甸，盘石等6个县46家养猪310个品种进行RT-PCR分析， PCR结果均为阳性，并将 PCR结果进行分析，并将 PCR结果与上海生工区进行比较。经检验，46家养猪中19家为 PEDV感染的养猪，其检出率达41.3%;310家养猪中，检出 PEDV的80家，其检出率为25.8%；永吉养猪的阳性率分别为58.33%和16.67%，桦甸地区为16.67%；在各季中，各地区的感染率以冬季最高。结果表明： PED发病率与各地区、各猪场免疫程序、季节和温度变化有密切的联系，且与我国大部分的病原有密切的联系。
关键词：猪流行性腹泻病毒 分子生物学 流行病学
Abstract
Porcine epidemic diarrhea (PED) is a highly infectious enteritis caused by porcine epidemic diarrhea disease (PEDV). The disease developed from the initial epidemic season (November to March of the next year) to a disease that can be epidemic once a year. The positive rate on the surface of pigs and the detection rate of meat products were 52.24% and 1.87%, respectively. The current epidemic situation of PED in Jilin cannot be effectively prevented. Therefore, there is a certain difference between PEDV infection in this region and that in China. In order to further understand the genetic variation and distribution characteristics of rural PEDV in Jilin Province, it has a good guiding significance for the prevention and control of rural PEDV in Jilin Province. At the same time, corresponding emergency treatment methods should be taken in time to minimize production costs and reduce the risk of production and operation.
In this paper, Primer 5.0 method was used to analyze 310 breeds of 46 pigs raised in Jilin City and Shulan, Jiaohe, Yongji, Huadian, Panshi and other six counties from February 2016 to February 2018 by using the PEDV gene sequence released by GenBank. The PCR results were all positive, and the PCR results were analyzed and compared with those in Shanghai production area. After inspection, 19 out of 46 pigs were infected with PEDV, and the detection rate was 41.3%; Among 310 pigs, 80 were found to have PEDV, with a detection rate of 25.8%; The positive rates of Yongji pigs were 58.33% and 16.67%, respectively, and 16.67% in Huadian area; In each season, the infection rate in each region was the highest in winter. The results showed that the incidence rate of PED was closely related to the immunization program, season and temperature changes in various regions and pig farms, and was closely related to most pathogens in China.
Key words: Porcine epidemic diarrhea virus Molecular Biology epidemiology
目 录
摘 要 I
Abstract II
1 猪流行性腹泻的研究进展 1
1.1 病原学 1
1.1.1 病原分类 1
1.1.2病原形态 1
1.1.3病毒基因组 1
1.1.4病毒理化特征 1
1.1.5病毒抗原性 1
1.1.6 病毒培养特性 2
1.2 流行病学 2
1.2.1 易感动物 2
1.2.2传播途径 2
1.2.3传染源 2
1.3流行动态 2
1.3.1 国外流行情况 3
1.4发病机理 4
1.5临床症状 4
1.5.1 哺乳仔猪 4
1.5.2断奶仔猪及保育仔猪 5
1.5.3生长育肥猪 5
1.5.4种母猪 5
1.5.5种公猪 5
1.6病理变化 5
1.6.1 眼观病理变化 5
1.6.2 镜观病理变化 5
1.7诊断方法 6
1.7.1 病原学诊断 7
1.7.2 血清学诊断 7
1.8 PED的防治 8
1.8.1 疫苗研究与应用 8
1.8.2 PED 的防控 10
2 吉林市猪流行性腹泻分子流行病学调查 10
2.1 试验材料 11
2.1.1 主要试剂 11
2.1.2 主要仪器 11
2.1.3 引物 12
2.1.4毒株 12
2.2 试验方法 12
2.2.1病料采集 12
2.2.2 RNA 提取 12
2.2.3 电泳 13
2.2.4凝胶回收 13
2.2.5克隆与测序 13
3 结果 14
3.1 PCR鉴定结果 14
3.2 吉林省规模化猪场不同季节仔猪病料 PCR 结果 15
3.3 测序结果 15
4 讨论 16
5 结论 17
参考文献 18
致 谢 20
