Preparation and evaluation of a recombinant Rift Valley fever virus N protein for the detection of IgG and IgM antibodies in humans and animals by indirect ELISA.
This paper describes the cloning, sequencing and bacterial expression of the N protein of the Rift Valley fever virus (RVFV) ZIM688/78 isolate and its evaluation in indirect ELISAs (I-ELISA) for the detection of IgM and IgG antibodies in human and sheep sera. Sera used for the evaluation were from 106 laboratory workers immunised with an inactivated RVF vaccine, 16 RVF patients, 168 serial bleeds from 8 sheep experimentally infected with wild type RVFV and 210 serial bleeds from 10 sheep vaccinated with the live attenuated Smithburn RVFV strain. All human and animal sera that tested positive in the virus neutralisation test were also positive in the IgG I-ELISA. There was a high correlation (R2=0.8571) between virus neutralising titres and IgG I-ELISA readings in human vaccinees. In experimentally infected sheep IgG antibodies were detected from day 4 to 5 post-infection onwards and IgM antibodies from day 3 to 4. The IgG I-ELISA was more sensitive than virus neutralisation and haemagglutination-inhibition tests in detecting the early immune response in experimentally infected sheep. The I-ELISAs demonstrated that the IgG and IgM response to the Smithburn vaccine strain was slower and the levels of antibodies induced markedly lower than to wild type RVFV infection.
Jansen van Vuren
Petrus
P
North-West University, Potchefstroom 2520, South Africa.
Potgieter
Abraham C
AC
Paweska
Janusz T
JT
van Dijk
Alberdina A
AA
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000662:veterinary; D005069:Evaluation Studies as Topic; D006386:Hemagglutination Tests; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D009500:Neutralization Tests; D021141:Nucleic Acid Amplification Techniques; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000276:immunology*; D012367:RNA, Viral / Q000032:analysis; D011994:Recombinant Proteins / Q000276:immunology; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012295:Rift Valley Fever / Q000276:immunology* / Q000662:veterinary; D012296:Rift Valley fever virus / Q000276:immunology*; D012680:Sensitivity and Specificity; D012756:Sheep; D013997:Time Factors; D014611:Vaccination
D000914:Antibodies, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D019590:Nucleocapsid Proteins; D012367:RNA, Viral; D011994:Recombinant Proteins