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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| Veröffentlicht in: | Journal of virological methods Jg. 63; H. 1; S. 47 - 56 |
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| Format: | Journal Article |
| Sprache: | Englisch |
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Elsevier B.V
1997
Amsterdam Elsevier New York, NY |
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| 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. |
| Author_xml | – sequence: 1 givenname: C.H. surname: Boshoff fullname: Boshoff, C.H. organization: Onderstepoort Veterinary Institute, Onderstepoort 0110, South Africa – sequence: 2 givenname: B. surname: Dungu fullname: Dungu, B. organization: Onderstepoort Veterinary Institute, Onderstepoort 0110, South Africa – sequence: 3 givenname: R. surname: Williams fullname: Williams, R. organization: Onderstepoort Veterinary Institute, Onderstepoort 0110, South Africa – sequence: 4 givenname: J. surname: Vorster fullname: Vorster, J. organization: Onderstepoort Veterinary Institute, Onderstepoort 0110, South Africa – sequence: 5 givenname: J.D. surname: Conradie fullname: Conradie, J.D. organization: Natal Institute of Immunology, Pinetown, Natal, South Africa – sequence: 6 givenname: D.W. surname: Verwoerd fullname: Verwoerd, D.W. organization: Onderstepoort Veterinary Institute, Onderstepoort 0110, South Africa – sequence: 7 givenname: D.F. surname: York fullname: York, D.F. organization: Department of Virology, Natal University School of Medicine, Durban 4000, South Africa |
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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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| 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 |
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