Development of a competitive ELISA for detecting antibodies to the peste des petits ruminants virus using a recombinant nucleoprotein. 1995

G Libeau, and C Préhaud, and R Lancelot, and F Colas, and L Guerre, and D H Bishop, and A Diallo
CIRAD-EMVT, Laboratoire de Pathotrop, Maisons-Alfort, France.

A competitive ELISA based on the reaction between a monoclonal antibody (mAb) and a recombinant nucleoprotein of the peste des petits ruminants virus (PPRV) was developed. This protein was obtained in large quantities from insect cells infected with a PPR nucleoprotein recombinant baculovirus (N-B). The competitive ELISA was compared with the virus neutralisation test (VNT) for detecting specific antibodies to PPRV in sheep and goats. The time consuming VNT is the only prescribed test that is capable of distinguishing between PPRV and the cross-reactive rinderpest virus (RPV). The competitive ELISA involves the simultaneous addition of the mAb and antibodies present in a positive serum, leading to competition for a specific epitope on the N-B. Optimum conditions were obtained by using serum samples which had positive or negative neutralising activity against PPRV or RPV. A negative cut-off point was determined on PPRV-negative sera from RPV-vaccinated cattle. A threshold value of 48 per cent inhibition, calculated from the mean for this population plus 2.7 standard deviations, was used in routine testing. A total of 683 sera were analysed by the competitive ELISA and the VNT. A good correlation (r = 0.94) was observed between the titres obtained in the two tests, with 80 sera that were from laboratory sources. The agreement between the two tests was determined on 271 field sera (kappa = 0.825). Their relative sensitivity (94.5 per cent) and specificity (99.4 per cent) were assessed on the 148 laboratory sera plus the 271 sera used for the determination of kappa.(ABSTRACT TRUNCATED AT 250 WORDS)

UI MeSH Term Description Entries
D009698 Nucleoproteins Proteins conjugated with nucleic acids. Nucleoprotein
D011994 Recombinant Proteins Proteins prepared by recombinant DNA technology. Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D004797 Enzyme-Linked Immunosorbent Assay An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed. ELISA,Assay, Enzyme-Linked Immunosorbent,Assays, Enzyme-Linked Immunosorbent,Enzyme Linked Immunosorbent Assay,Enzyme-Linked Immunosorbent Assays,Immunosorbent Assay, Enzyme-Linked,Immunosorbent Assays, Enzyme-Linked
D006041 Goats Any of numerous agile, hollow-horned RUMINANTS of the genus Capra, in the family Bovidae, closely related to the SHEEP. Capra,Capras,Goat
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D000911 Antibodies, Monoclonal Antibodies produced by a single clone of cells. Monoclonal Antibodies,Monoclonal Antibody,Antibody, Monoclonal
D000914 Antibodies, Viral Immunoglobulins produced in response to VIRAL ANTIGENS. Viral Antibodies
D012756 Sheep Any of the ruminant mammals with curved horns in the genus Ovis, family Bovidae. They possess lachrymal grooves and interdigital glands, which are absent in GOATS. Ovis,Sheep, Dall,Dall Sheep,Ovis dalli
D012757 Sheep Diseases Diseases of domestic and mountain sheep of the genus Ovis. Ovine Diseases,Disease, Ovine,Disease, Sheep,Diseases, Ovine,Diseases, Sheep,Ovine Disease,Sheep Disease
D014764 Viral Proteins Proteins found in any species of virus. Gene Products, Viral,Viral Gene Products,Viral Gene Proteins,Viral Protein,Protein, Viral,Proteins, Viral

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