猪流行性腹泻胶体金抗体检测技术的建立及其应用 - 中国农学通报.pdf

中国农学通报 2011,27(03):374-377 Chinese Agricultural Science Bulletin 猪流行性腹泻胶体金抗体检测技术的建立及其应用 张利勃，周铁忠，王 珅，高慎阳 （辽宁医学院畜牧兽医学院重点实验室，辽宁锦州121001） 摘 要：为建立一种高效、经济、简便适用于基层防疫检疫人员使用的猪流行性腹泻(PED)疾病的检测方 法，参考已知的胶体金免疫层析法(GICA)工作原理基础之上，将葡萄球菌蛋白A(SPA)进胶体金标记作 为指示介质，将基因工程表达PEDV M 蛋白抗原和自制的抗SPA 多抗血清包被硝酸纤维素膜(NCM) 分 别作为检测线和质控线，由此制成一种检测PEDV 血清抗体的快速诊断试纸。其特异性、敏感性试验与 ELISA 进行比较。结果表明该GICA 与参考检测方法ELISA 具有相近的灵敏度，在二者检测的75 份临 床样本中有65 份共同检测为阳性结果，7 份共同检测为阴性结果，其符合率达96% 。说明该GICA 检测 技术的建立为检测PEDV 抗体提供了一种快速简便的方法。 关键词：猪流行性腹泻；胶体金免疫层析测定；酶联免疫吸附测定；葡萄球菌蛋白A 中图分类号：S855.3 文献标志码：A 论文编号：2010-2557 DevelopmentandPreliminaryApplicationofaGold-ImmunochromatographyAssayforPED Zang Libo, Zhou Tiezhong, Wang Shen, Gao Shenyang (KeyLaboratoryofAnimalHusbandryandVeterinaryMedicine, LiaoningMedicalUniversity, Jinzhou Liaoning 121001) Abstract: To establish a highly efficient, cost-effective and handily detection method for the diagnosis of porcine epidemic diarrhea. Based on the well known working principle of Gold-Immunochromatography Assay (GICA), a batch of in-house GICA test strips were made by assembling the following elements, which were included the protein SPA labeled with HAuCl4 as indicating reagent as well as the recombinant PEDV M protein and multiple anti-SPA serum coated onto the NC membrane as detective line and control line respectively. Compared with the reference ELISA method, the established GICA shared the similar test sensitivity. Especially, the clinical test agreement rate of both methods reached 96%, which was calculated by the sum percent of positive (65) and negative (7) samples among total 75 clinical samples. This GICA method provided the most rapid and convenient means for detection of PED