猪流行性腹泻病毒抗体间接ELISA检测方法的建立与应用
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广东省科技计划项目(2011B090400414); 广州市科技计划项目(201314100051)


Development and application of indirect ELISA for porcine epidemic diarrhea virus antibody detection
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    【目的】建立2种猪流行性腹泻病毒(PEDV)抗体间接ELISA检测方法.【方法】分别以GD-1株PEDV灭活全病毒和原核表达的COE融合蛋白作为包被抗原,确定最佳ELISA条件,检验其重复性、特异性等,并进行临床应用.【结果和结论】确定了2种ELISA方法的最佳条件,重复性试验显示变异系数分别小于4.60%和5.30%,特异性试验检测口蹄疫病毒、猪瘟病毒、猪圆环病毒和猪繁殖与呼吸综合征病毒阳性血清均为阴性,并具有良好的稳定性和较长的保存期.临床样品检测结果显示,223份血清样品总阳性率分别为97.3% (217/223)和98.6% (220/223),2种方法检测的阳性率相近,具有良好的检出率和很高的符合率,为PEDV的抗体检测及流行病学调查提供了一种技术手段.

    Abstract:

    【Objective】Two types of ELISA methods for porcine epidemic diarrhea virus (PEDV) antibody detection were established.【Method】 The inactivated PEDV strain GD-1 and expressed COE protein as coating antigen were used to establish optimal ELISA conditions and test the repeatability, specificity and clinical application effectiveness.【Result and conclusion】 The optimal conditions of two types of ELISA were established. The variation coefficient of two EILSA methods was less than 4.60% and 5.30% respectively. The specific test results of foot and mouth disease virus(FMDV), classical swine fever virus(CSFV), porcine circovirus(PCV), porcine reproductive and respiratory syndrome virus(PRRSV) positive serum were all negative. The total positive rate of 223 clinical serum samples was 97.3% (217/223) and 98.6% (220/223) respectively. These two methods have a good stability and long shelf life. This research proves that two types of ELISA methods have a good detection rate and a high coincidence rate which will provide a technical method for PEDV antibody detection and epidemiological investigation.

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曾喜多,魏钟艳,李智丽,孙宝丽,谢青梅,毕英佐,马静云.猪流行性腹泻病毒抗体间接ELISA检测方法的建立与应用[J].华南农业大学学报,2015,36(4):16-20

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  • 收稿日期:2014-06-30
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  • 在线发布日期: 2015-07-10