贝类帕金虫间接酶联免疫吸附测定法的建立  

Establishment of indirect enzyme-linked immunosorbent assay(iELISA) of detecting Perkinsus marinus in mollusc

作　　者：樊景凤[1] 梁玉波[1] 宋立超[2] 张喜昌[3] 

机构地区：[1]国家海洋环境监测中心,辽宁大连116023 [2]沈阳农业大学土地与环境学院,辽宁沈阳110161 [3]中国科学院大连化学物理研究所,辽宁大连116023

出　　处：《海洋环境科学》2006年第2期62-64,共3页Marine Environmental Science

基　　金：国家863高技术研究发展计划项目(2002AA639470);国家海洋局重点青年基金项目(2001303)

摘　　要：采用液体巯基醋酸盐培养基(FTM)培养贝类帕金虫,以其制备抗帕金虫的免疫血清,建立一种快速检测贝类帕金虫的间接酶联免疫吸附测定法(iELISA)。方法中理想的包被抗原数量为104/mL;帕金虫抗血清最佳工作浓度1:10 000;酶标二抗工作浓度为1:1 000;进行血清敏感性测定,其最低检出限为102/mL;阻断试验中的阻断率达86.77%;板内和板间变异系数分别为2.9%和3.21%。将该方法标准化后,对30份菲律宾蛤仔体内帕金虫进行检测,所得结果与用FTM方法得到的结果进行比较,后者灵敏度要高于前者,但是在检测时间上,前者在24 h内能够完成,而后者需要7 d。An indirect enzyme-linked immunosorbent assay （iEL1SA） for rapid diagnosis of Perkinsus marinus in mollusc has been developed. The Perkinsus marinus were cultured by fluid thioglycollate medium（ FTM ）. In order to gain the polyclonal antibody against Perkinsus marinus,the Perkinsus marinus immuned rabbits. The optimal concentration of the coating antigen was 104/mL, the dilutions of polyclonal antibody against Perkinsus marinus and goat anti-rabbit lgG were 1/10 000 and 1/1 000 respectively. The sensitivity of the serum was tested,and the lowest Perkinsus marinus suspension was 102/mL. Inhibition rate was 86.77% in inhibiton test. The coefficients of variation of intra-assay and inter assay were 2.9% and 3.21% respectively. The method was optimized to detect the Perkinsus marinus in Ruditapes philippinarum, and to compare with FTM ： the results indicated the latter was more sensitive than the former, howerver, the former were accomplished in 24 hours and the latter need 7 days.

关 键 词：贝类帕金虫 间接ELISA 检测 

分 类 号：S944.4[农业科学—水产养殖]