P49重组抗原检测旋毛虫抗体的间接ELISA方法建立  

Establishment of an indirect ELISA for the detection of antibodies against P49 recombinant protein of Trichinella nativa

作　　者：李晓云[1] 路义鑫[1] 韩彩霞[1] 朱江巍[1] 宋铭忻[1] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《中国兽医杂志》2011年第12期7-9,共3页Chinese Journal of Veterinary Medicine

基　　金："十一五"国家科技支撑计划重大项目(2010BAD04B01);家畜疫病病原生物学国家重点实验室开放课题(SKLVEB2009KFKT005);黑龙江省普通高等学校青年学术骨干支持计划项目(1153G007)

摘　　要：以纯化的本地毛形线虫P49排泄分泌(ES)重组蛋白包被微量反应板,建立检测旋毛虫抗体的间接ELISA方法,并确定了最适反应参数:抗原包被浓度4μg/mL;血清稀释度1∶100,作用时间90min;酶标二抗稀释度1∶10 000,作用时间60min;最适封闭液为5g/L聚乙烯醇(PVA);最适稀释液为含20g/L PEG6000的PBS;底物最佳显色时间15min。ELISA体系评价结果表明,该方法重复性好,灵敏度高,且可用于不同种株旋毛虫感染的P49血清抗体检测。An indirect ELISA was established for the detection of antibodies against Trichinella nativa using P49 excretory-secretory （ES） recombinant protein as coating antigen. In the ELISA, the optimal concentration of P49 was 4 μg/mL, and serum should be diluted 200 times and incubated with the antigen at 37℃ for 90 min, the secondary antibody should be diluted at 1.10 000 times and incubated with the serum at 37℃ for 60 min, the optimal blocking solution was 5 g/L PVA and the optimal PEG6000 concentration in dilution was 20 g/L, and the substrate should be incubated for 15 min. Evaluation results of the ELISA system showed that the method had good reproducibility and sensitivity, and was suitable to detect antiserum against P49 protein of different Trichinella spp.

关 键 词：本地毛形线虫 P49重组抗原 间接ELISA 

分 类 号：S852.731[农业科学—基础兽医学]