title: Development of indirect ELISAs for differential serodiagnosis of classical and highly pathogenic porcine reproductive and respiratory syndrome virus.
abstract: 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 OD(450nm) for the iELISA-180 and 0.27 at OD(450nm) 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.
authors.affiliation: Department of Basic Veterinary Medicine, College of Veterinary Medicine, Shandong Agricultural University, Shandong, China.
authors.forename: Y H
authors.initials: YH
authors.lastname: Xiao
authors.forename: T T
authors.initials: TT
authors.lastname: Wang
authors.forename: Q
authors.initials: Q
authors.lastname: Zhao
authors.forename: C B
authors.initials: CB
authors.lastname: Wang
authors.forename: J H
authors.initials: JH
authors.lastname: Lv
authors.forename: L
authors.initials: L
authors.lastname: Nie
authors.forename: J M
authors.initials: JM
authors.lastname: Gao
authors.forename: X C
authors.initials: XC
authors.lastname: Ma
authors.forename: W H
authors.initials: WH
authors.lastname: Hsu
authors.forename: E M
authors.initials: EM
authors.lastname: Zhou
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000276:immunology*; D015153:Blotting, Western; D003937:Diagnosis, Differential; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D019318:Porcine Reproductive and Respiratory Syndrome / Q000097:blood / Q000175:diagnosis* / Q000517:prevention & control / Q000821:virology; D019316:Porcine respiratory and reproductive syndrome virus / Q000276:immunology* / Q000302:isolation & purification / Q000472:pathogenicity; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D013552:Swine; D014765:Viral Vaccines / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D011994:Recombinant Proteins; D014765:Viral Vaccines