Development of Indirect ELISAs for Differential Serodiagnosis of Classical and Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus
Development of Indirect ELISAs for Differential Serodiagnosis of Classical and Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus
复制标题
开发用于经典和高致病性猪繁殖与呼吸综合征病毒血清学鉴别诊断的间接 ELISA
DOI：
10.1111/tbed.12040
复制
发表时间：
2014-08-01
影响因子：
4.3
通讯作者：
Zhou, E. M.
中科院分区：
文献类型：
--
作者：
Xiao, Y. H.;Wang, T. T.;Zhou, E. M.
本研究建立了两种检测猪繁殖与呼吸综合征病毒(PRRSV)和高致病性PRRSV(HP-PRRSV)疫苗株血清抗体的间接酶联免疫吸附试验(IELISA)。为了检测常见的抗经典和HP-PRRSV抗体，间接酶联免疫吸附试验(iEL ISA-180)所用的包被抗原是Nsp2-180的抗原，Nsp2的氨基末端的180aa。用包被抗原Nsp2-D29作为包被抗原，检测Hp-PRRSV与经典和Hp-PRRSV不同的抗体。采用纸板滴定优化了抗原浓度和血清稀释度。通过对120份PRRSV阳性和198份PRRSV阴性猪血清的检测，确定了iEL ISA-180和iEL ISA-D29的诊断界值分别为0.361和0.27，其特异度分别为97.1%和96.7%，敏感性分别为96.9%和96.3%。Western印迹与iEL ISA-180、iEL ISA-D29的符合率分别为98%、96.7%。该方法可用于临床血清标本中HP-PRRSV与疫苗或经典PRRSV的血清学鉴别。
The objective of this study was to develop two indirect enzyme-linked immunosorbent assays (iELISAs) for detection of serum antibodies against classical vaccine strain of porcine reproductive and respiratory syndrome virus (PRRSV) and highly pathogenic PRRSV (HP-PRRSV). To detect the common antibodies against classical and HP-PRRSV, the coating antigen used in the iELISA (designated iELISA-180) was the antigen of Nsp2-180, the 180aa at amino terminal of Nsp2. To detect the different antibodies against classical and HP-PRRSV, the coating antigen in the second iELISA (designated iELISA-D29) was Nsp2-D29, the deleted 29aa in Nsp2 of HP-PRRSV. The antigen concentration and serum dilutions were optimized using a draughtboard titration. The cut-off values of 0.361 at OD450nm for the iELISA-180 and 0.27 at OD450nm for the iELISA-D29 were determined by testing a panel of 120 classical PRRSV positive and 198 PRRSV negative pig serum samples, which generated the specificity of 97.1% and 96.7%, the sensitivity of 96.9% and 96.3% for iELISA-180 and iELISA-D29, respectively. The agreements between the Western blot and iELISA-180 and iELISA-D29 were 98%, 96.7%, respectively. The developed iELISAs can be used to differentiate serologically HP-PRRSV from the vaccinated or classical PRRSV in clinical serum samples.