|
|||||
|
|
| 猪流行性腹泻病毒间接ELISA检测方法的建立 | |
| 引用本文： | 王丽珍,罗桂芳,张艳萍,沈勇,吴源潇,石燕,陈瑞爱,贺东生.猪流行性腹泻病毒间接ELISA检测方法的建立[J].中国畜牧兽医,2017,44(2):377-383. |
| 作者姓名： | 王丽珍 罗桂芳 张艳萍 沈勇 吴源潇 石燕 陈瑞爱 贺东生 |
| 作者单位： | 1. 华南农业大学兽医学院, 广州 510642; 2. 广东温氏大华农生物科技有限公司, 广州 510642 |
| 基金项目： | 大学生创新创业训练计划项目（201510564099） |
| 摘 要： | 猪流行性腹泻（porcine epidemic diarrhea，PED）是由猪流行性腹泻病毒（porcine epidemic diarrhea virus，PEDV）引起的猪的一种急性、高度传染性肠道疾病，给养猪业造成了严重的经济损失。本研究以纯化的PEDV 为包被抗原，通过优化 ELISA 反应条件，建立了间接 ELISA 抗体检测方法，其反应条件为：抗原最佳包被浓度为 20 μg/mL，血清标准品最佳稀释度为 1：500，包被时间为4℃过夜，5%小牛血清37℃封闭1 h，二抗 1：10 000稀释，37℃作用1 h，抗体临界值为 D450 nm≥ 0.289判为阳性，D450 nm≤ 0.236判为阴性，介于二者之间为可疑。检测猪繁殖与呼吸综合征病毒、猪圆环病毒2型、猪瘟病毒、伪狂犬病毒、细小病毒和口蹄疫病毒标准血清抗体均为阴性，重复试验变异系数均小于10%，对江苏、江西、福建、广东地区74份猪血清样品进行检测，抗体阳性率达84%，表明该方法具有较好敏感性、特异性和重复性，可用于 PEDV 抗体检测和流行病学调查。 |
| 关 键 词： | 猪流行性腹泻病毒 ELISA 包被 抗体 |
| 收稿时间： | 2016-07-12 |
Establishment of Indirect ELISA Detection Method for Porcine Epidemic Diarrhea Virus |
|
| WANG Li-zhen,LUO Gui-fang,ZHANG Yan-ping,SHEN Yong,WU Yuan-xiao,SHI Yan,CHEN Rui-ai,HE Dong-sheng.Establishment of Indirect ELISA Detection Method for Porcine Epidemic Diarrhea Virus[J].China Animal Husbandry & Veterinary Medicine,2017,44(2):377-383. | |
| Authors: | WANG Li-zhen LUO Gui-fang ZHANG Yan-ping SHEN Yong WU Yuan-xiao SHI Yan CHEN Rui-ai HE Dong-sheng |
| Institution: | 1. College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China; 2. Guangdong Wens Dahuanong Biotechnology Co., Ltd., Guangzhou 510642, China |
| Abstract: | Porcine epidemic diarrhea (PED) is an acute, highly contagious enteric disease of pigs. Porcine epidemic diarrhea virus (PEDV) is the causative agent of PED. PED has caused significant economic losses to the pig industry. In this study,the purified PEDV as the coating antigen, by optimizing the ELISA reaction conditions,the indirect ELISA antibody detection method was established. The optimized reaction conditions were as follows: Antigen working concentration was 20 μg/mL; Serum sample dilution was 1:500;It was coated at 4℃ overnight; The plates were blocked by 5% calf serum incubated at 37℃ for 1 h; The secondary antibody was diluted at 1:10 000,incubated at 37℃ for 1 h. It was judged as positive when the cutoff value D450 nm≥0.289,as negative when D450 nm≤0.236,and as suspicious between 0.289 and 0.236.It could not react with the positive sear of other six viruses such as porcine respiratory and reproductive syndrome virus,porcine circovirus 2, classical swine fever virus, porcine parvovirus,pseudo rabies virus and foot-and-mouth disease virus. The variation coefficient of repeated test was less than 10%.74 pig serum samples from Jiangsu, Jiangxi, Fujian and Guangdong were detected,and the positive rate was 84%.It indicated that this method could be used for PEDV epidemiological surveys and diagnosis in the future. |
| Keywords: | porcine epidemic diarrhea virus (PEDV) ELISA coat antibody |
| 本文献已被 CNKI 等数据库收录！ | |
| 点击此处可从《中国畜牧兽医》浏览原始摘要信息 | |
| 点击此处可从《中国畜牧兽医》下载免费的PDF全文 | |