Detection of Maedi-Visna virus antibodies using a single fusion transmembrane-core p25 recombinant protein ELISA and a modified receiver-operating characteristic analysis to determine cut-off values

The core p25 and transmembrane (TM) genes of Maedi-Visna virus (MVV) were cloned individually into the pGEX-2T expression vector. Both proteins were expressed as a combined fusion protein in frame with glutathione S-transferase (GST). The purified recombinant antigens (GST-TM and GST-TM-p25) were us...

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Vydáno v:	Journal of virological methods Ročník 63; číslo 1; s. 47 - 56
Hlavní autoři:	Boshoff, C.H., Dungu, B., Williams, R., Vorster, J., Conradie, J.D., Verwoerd, D.W., York, D.F.
Médium:	Journal Article
Jazyk:	angličtina
Vydáno:	London Elsevier B.V 1997 Amsterdam Elsevier New York, NY
Témata:	Animal viral diseases Animals Antibodies, Viral - blood Antibodies, Viral - immunology Antigens, Viral - genetics Antigens, Viral - immunology Biological and medical sciences Cloning, Molecular ELISA Enzyme-Linked Immunosorbent Assay - methods Escherichia coli Gene Products, gag - genetics Gene Products, gag - immunology Infectious diseases Maedi-Visna virus diagnosis Medical sciences Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - immunology Sheep Two-graph receiver operating characteristic Viral diseases Visna - blood Visna - immunology Visna - virology Visna-maedi virus - immunology Visna-maedi virus - isolation & purification Maedi-Visna virus diagnosis Two-graph receiver operating characteristic ELISA Transmembrane protein Antibody Retroviridae Visna virus Infection Virus Vertebrata Mammalia Viral disease Animal Serological method Lentivirinae Sheep Artiodactyla Diagnosis Recombinant protein Veterinary Detection Maedi ELISA assay Ungulata Visna Maedi virus
ISSN:	0166-0934, 1879-0984
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Abstract	The core p25 and transmembrane (TM) genes of Maedi-Visna virus (MVV) were cloned individually into the pGEX-2T expression vector. Both proteins were expressed as a combined fusion protein in frame with glutathione S-transferase (GST). The purified recombinant antigens (GST-TM and GST-TM-p25) were used to develop a MVV ELISA. A preliminary assessment of the diagnostic potential of the recombinant antigens (GST-TM and GST-TM-p25) was made by testing the antigens against 46 seropositive and 46 seronegative sheep and comparing the results with a commercial p25 ELISA kit. A two-graph receiver operating characteristic (TG-ROC) analysis program was used to interpret the data. The GST-TM-p25 ELISA was more sensitive than the commercial assay which is based on the p25 antigen alone and more specific than the GST-TM ELISA.
AbstractList	The core p25 and transmembrane (TM) genes of Maedi-Visna virus (MVV) were cloned individually into the pGEX-2T expression vector. Both proteins were expressed as a combined fusion protein in frame with glutathione S-transferase (GST). The purified recombinant antigens (GST-TM and GST-TM-p25) were used to develop a MVV ELISA. A preliminary assessment of the diagnostic potential of the recombinant antigens (GST-TM and GST-TM-p25) was made by testing the antigens against 46 seropositive and 46 seronegative sheep and comparing the results with a commercial p25 ELISA kit. A two-graph receiver operating characteristic (TG-ROC) analysis program was used to interpret the data. The GST-TM-p25 ELISA was more sensitive than the commercial assay which is based on the p25 antigen alone and more specific than the GST-TM ELISA. The core p25 and transmembrane (TM) genes of Maedi-Visna virus (MVV) were cloned individually into the pGEX-2T expression vector. Both proteins were expressed as a combined fusion protein in frame with glutathione S-transferase (GST). The purified recombinant antigens (GST-TM and GST-TM-p25) were used to develop a MVV ELISA. A preliminary assessment of the diagnostic potential of the recombinant antigens (GST-TM and GST-TM-p25) was made by testing the antigens against 46 seropositive and 46 seronegative sheep and comparing the results with a commercial p25 ELISA kit. A two-graph receiver operating characteristic (TG-ROC) analysis program was used to interpret the data. The GST-TM-p25 ELISA was more sensitive than the commercial assay which is based on the p25 antigen alone and more specific than the GST-TM ELISA. The core p25 and transmembrane (TM) genes of Maedi-Visna virus (MVV) were cloned individually into the pGEX-2T expression vector. Both proteins were expressed as a combined fusion protein in frame with glutathione S-transferase (GST). The purified recombinant antigens (GST-TM and GST-TM-p25) were used to develop a MVV ELISA. A preliminary assessment of the diagnostic potential of the recombinant antigens (GST-TM and GST-TM-p25) was made by testing the antigens against 46 seropositive and 46 seronegative sheep and comparing the results with a commercial p25 ELISA kit. A two-graph receiver operating characteristic (TG-ROC) analysis program was used to interpret the data. The GST-TM-p25 ELISA was more sensitive than the commercial assay which is based on the p25 antigen alone and more specific than the GST-TM ELISA.The core p25 and transmembrane (TM) genes of Maedi-Visna virus (MVV) were cloned individually into the pGEX-2T expression vector. Both proteins were expressed as a combined fusion protein in frame with glutathione S-transferase (GST). The purified recombinant antigens (GST-TM and GST-TM-p25) were used to develop a MVV ELISA. A preliminary assessment of the diagnostic potential of the recombinant antigens (GST-TM and GST-TM-p25) was made by testing the antigens against 46 seropositive and 46 seronegative sheep and comparing the results with a commercial p25 ELISA kit. A two-graph receiver operating characteristic (TG-ROC) analysis program was used to interpret the data. The GST-TM-p25 ELISA was more sensitive than the commercial assay which is based on the p25 antigen alone and more specific than the GST-TM ELISA.
Author	Conradie, J.D. Verwoerd, D.W. Boshoff, C.H. Dungu, B. Williams, R. York, D.F. Vorster, J.
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Keywords	Maedi-Visna virus diagnosis Two-graph receiver operating characteristic ELISA Transmembrane protein Antibody Retroviridae Visna virus Infection Virus Vertebrata Mammalia Viral disease Animal Serological method Lentivirinae Sheep Artiodactyla Diagnosis Recombinant protein Veterinary Detection Maedi ELISA assay Ungulata Visna Maedi virus
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Snippet	The core p25 and transmembrane (TM) genes of Maedi-Visna virus (MVV) were cloned individually into the pGEX-2T expression vector. Both proteins were expressed...
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SubjectTerms	Animal viral diseases Animals Antibodies, Viral - blood Antibodies, Viral - immunology Antigens, Viral - genetics Antigens, Viral - immunology Biological and medical sciences Cloning, Molecular ELISA Enzyme-Linked Immunosorbent Assay - methods Escherichia coli Gene Products, gag - genetics Gene Products, gag - immunology Infectious diseases Maedi-Visna virus diagnosis Medical sciences Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - immunology Sheep Two-graph receiver operating characteristic Viral diseases Visna - blood Visna - immunology Visna - virology Visna-maedi virus - immunology Visna-maedi virus - isolation & purification
Title	Detection of Maedi-Visna virus antibodies using a single fusion transmembrane-core p25 recombinant protein ELISA and a modified receiver-operating characteristic analysis to determine cut-off values
URI	https://dx.doi.org/10.1016/S0166-0934(96)02114-3 https://www.ncbi.nlm.nih.gov/pubmed/9015275 https://www.proquest.com/docview/78791267
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