title: Validation of an IgM antibody capture ELISA based on a recombinant nucleoprotein for identification of domestic ruminants infected with Rift Valley fever virus.
abstract: The presence of competent vectors in some countries currently free of Rift Valley fever (RVF) and global changes in climate, travel and trade have increased the risk of RVF spreading to new regions and have emphasised the need for accurate and reliable diagnostic tools for early diagnosis during RVF outbreaks. Highly sensitive viral detection systems like PCR have a limited use during outbreaks because of the short duration of viraemia, whereas antibodies like specific IgM which are serological indicators of acute infection, can be detected for up to 50 days after infection. Using the highly conserved and immunogenic recombinant nucleoprotein of RVF virus in an IgM capture ELISA, the risk of laboratory infection associated with traditional serological methods is avoided. The use of pre-coated/pre-blocked ELISA plates and the conjugation of the recombinant nucleoprotein with horseradish peroxidase simplified and shortened the assay procedure. Results showed the assay to be highly reproducible with a lower detection limit equal to that of a commercial competition ELISA. By receiver operating characteristic (ROC) curve analysis the area under curve (AUC) index was determined as 1.0 and the diagnostic sensitivity and specificity at a PP cut-off value of 4.1 as 100% and 99.78% respectively. The results of this study demonstrated that the IgM capture ELISA is a safe, reliable and highly accurate diagnostic tool which can be used on its own or in parallel with other methods for the early diagnosis of RVF virus infection and also for monitoring of immune responses in vaccinated domestic ruminants.
authors.affiliation: ARC-Onderstepoort Veterinary Institute, Private Bag X05, Onderstepoort, Gauteng, 0110, South Africa. WilliamsR@arc.agric.za
authors.forename: Roy
authors.initials: R
authors.lastname: Williams
authors.forename: Charlotte Elizabeth
authors.initials: CE
authors.lastname: Ellis
authors.forename: Shirley Jacqueline
authors.initials: SJ
authors.lastname: Smith
authors.forename: Christiaan Abraham
authors.initials: CA
authors.lastname: Potgieter
authors.forename: David
authors.initials: D
authors.lastname: Wallace
authors.forename: Vuyokazi Epipodia
authors.initials: VE
authors.lastname: Mareledwane
authors.forename: Phelix Antipas Ochola
authors.initials: PA
authors.lastname: Majiwa
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D019540:Area Under Curve; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000662:veterinary; D007106:Immune Sera / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology; D058751:Livestock / Q000821:virology; D019590:Nucleocapsid Proteins / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D015203:Reproducibility of Results; D012295:Rift Valley Fever / Q000175:diagnosis* / Q000276:immunology / Q000662:veterinary / Q000821:virology; D012296:Rift Valley fever virus / Q000276:immunology / Q000302:isolation & purification* / Q000472:pathogenicity; D012418:Ruminants / Q000276:immunology / Q000821:virology*; D012680:Sensitivity and Specificity
chemical_list: D000914:Antibodies, Viral; D007106:Immune Sera; D007075:Immunoglobulin M; D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins