中国动物传染病学报Issue(6)：12-18,7.
猪繁殖与呼吸综合征基因标记疫苗株NP49-ELISA鉴别诊断方法的建立
DEVELOPMENT OF NP49-ELISA FOR DIFFERENTIAL DIAGNOSIS FOR MARKER VACCINE AGAINST PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME
摘要
Abstract
Recombinant virus rHN4-Δ25+NP49 strain is a newly developed genetic marker vaccine against highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV). The marker gene NP49 represents 49 amino acids at C terminal of Newcastle disease virus NP protein. Reasonably, detection of the antibodies specific for marker gene NP49 would differentiate vaccinated pigs from field virus infected pigs. To develop a differental diagnosis, synthesized NP49 peptide was used as coating antigen in indirect ELISA. Then, the differential NP49-ELISA was optimized for coating antigen concentration, serum dilution, etc. The optimal coating concentration of NP49 antigen was 500 ng/well and optimal serum dilution was 1:40. The critical S/P value was determined at 0.2 according to the ROC curve. The intra-and inter-assay coefficients of variability between experiments were all less than 10%. There was no cross reaction with antibodies to other swine viruses, indicating good reproducibility and specificity. Results have demonstrated that NP49-ELISA is a reproducible and specific method for differential diagnosis of vaccinated pigs, paving a pathway for clinical application of PRRSV genetic marker vaccine rHN4-Δ25+NP49 strain.关键词
猪繁殖与呼吸综合征标记疫苗/多肽抗原/NP49-ELISA/鉴别诊断Key words
Porcine reproductive and respiratory syndrome, rirus/marker vaccine/peptide antigen/NP49-ELISA/differential diagnosis分类
农业科技引用本文复制引用
孙晶,周艳君,徐彦召,姜一峰,童武,童光志..猪繁殖与呼吸综合征基因标记疫苗株NP49-ELISA鉴别诊断方法的建立[J].中国动物传染病学报,2013,(6):12-18,7.基金项目
国家“863”计划项目（2011AA10A208-2）；国际合作项目（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）；国家自然科学基金项目（31100121）；中国农业科学院基本科研业务费项目 （）
