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

J Virol Methods. 1997 Jan;63(1-2):47-56. doi: 10.1016/s0166-0934(96)02114-3.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Viral / blood*
  • Antibodies, Viral / immunology
  • Antigens, Viral / genetics
  • Antigens, Viral / immunology*
  • Cloning, Molecular
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Escherichia coli
  • Gene Products, gag / genetics
  • Gene Products, gag / immunology*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Sheep
  • Visna / blood
  • Visna / immunology
  • Visna / virology*
  • Visna-maedi virus / immunology
  • Visna-maedi virus / isolation & purification*

Substances

  • Antibodies, Viral
  • Antigens, Viral
  • Gene Products, gag
  • Recombinant Fusion Proteins