Detection of antibodies to caprine arthritis-encephalitis virus using recombinant gag proteins

E. Rimstad, Nancy East, E. DeRock, J. Higgins, Niels C Pedersen

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

The coding sequences of the core proteins p17 and p28 of caprine arthritis-encephalitis virus (CAEV) were amplified using the polymerase chain reaction and cloned into the plasmid expression vector p-GEX-2T. Both p17 and p28 were expressed as fusion proteins with glutathione S-transferase. The recombinant proteins were affinity purified from induced bacterial lysates using glutathione-agarose beads. The purified proteins were used in an enzyme-linked immunosorbent assay (ELISA) to detect antibodies against CAEV in goat sera and milk samples. Three different ELISA tests were developed based on p17, p28 or the combination of these two recombinant proteins (p17+p28). A comparison was made to an ELISA based on purified whole virus particles and to agar immunodiffusion test (AGID). Sera with conflicting results in the different ELISA tests were examined by Western blotting. There was a high correlation between the ELISA tests based on p17+p28 recombinant proteins and whole virus ELISA, with an estimated κ value of 0.92. Only 72-75% of the sera that tested positive in these two ELISA tests were positive in AGID. Antibodies to CAEV were detected in significantly more animals when serum samples were tested compared to milk samples. Based on the time and materials required to prepare the reagents, the recombinant based ELISA test was less expensive than the whole virus ELISA.

Original languageEnglish (US)
Pages (from-to)345-356
Number of pages12
JournalArchives of Virology
Volume134
Issue number3-4
DOIs
StatePublished - Sep 1994

Fingerprint

Caprine Arthritis-Encephalitis Viruses
gag Gene Products
Recombinant Proteins
Enzyme-Linked Immunosorbent Assay
Antibodies
Immunodiffusion
Serum
Agar
Milk
Viruses
Proteins
Glutathione Transferase
Goats
Virion
Sepharose
Glutathione
Plasmids
Western Blotting

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Genetics

Cite this

Detection of antibodies to caprine arthritis-encephalitis virus using recombinant gag proteins. / Rimstad, E.; East, Nancy; DeRock, E.; Higgins, J.; Pedersen, Niels C.

In: Archives of Virology, Vol. 134, No. 3-4, 09.1994, p. 345-356.

Research output: Contribution to journalArticle

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