中国动物传染病学报Issue(1)：30-36,7.
高致病性猪繁殖与呼吸综合征病毒基因缺失标记疫苗ELISA鉴别诊断方法的建立
DEVELOPMENT OF ELISA DIFFERENTIATION DIAGNOSIS FOR A DUAL-MARKER VACCINE OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS
摘要
Abstract
To develop a ELISA method for detection of antibody against 25aa in Nsp2 region which was deleted from a dual-marker PRRSV vaccine strain rHN4-Δ25+NP49. The 25aa peptide was synthesized artificially and used as antigen to coat the microplate for ELISA. The optimal coating concentration of antigen was 500 ng/well. optimal serum dilution was 1:40, and the cut off S/P value was 0.15. The reproducibility test showed that the coefficients of variation for intra-and inter-assay were lower than 10%. 25aa-ELISA method was used to test clinical samples and compared with commercial PRRSV-detection-kit (IDEXX). The results showed that coincidence of two methods was 94.84%. Then 25aa-ELISA was used to test HuN4-F112 immunized serum samples from 1 to 126 days post immunization (dpi). Specific antibody against 25aa was detected during 21 to 126 dpi. The vaccine immunized pigs could be differentiated from naturally infected pigs with wild type PRRSVs. Therefore, this study set up a foundation for further investigation of the differentiation diagnosis for a dual-marker vaccine rHN4-Δ25+NP49.关键词
高致病性猪繁殖与呼吸综合征病毒/多肽抗原/25aa-ELISAKey words
Highly pathogenic Porcine reproductive and respiratory syndrome virus/peptide antigen/25aa-ELISA分类
农业科技引用本文复制引用
孙晶,周艳君,姜一峰,徐彦召,童武,童光志..高致病性猪繁殖与呼吸综合征病毒基因缺失标记疫苗ELISA鉴别诊断方法的建立[J].中国动物传染病学报,2014,(1):30-36,7.基金项目
国家863计划项目(2011AA10A208-2)；国家国际科技合作重点计划项目(2010DF33920)；科研院所技术开发研究专项资金(2012EG134237)；上海市科技兴农重点攻关项目(沪农科攻字(2012)第2-5号)；中国农业科学院基本科研业务费预算增量项目(2013BL039) （2011AA10A208-2）
