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M9610041.TXT
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1996-01-30
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Document 0041
DOCN M9610041
TI Bacterial expression of the caprine arthritis-encephalitis virus gag and
env proteins and their use in enzyme-linked immunosorbent assay.
DT 9601
AU Clavijo A; Thorsen J; Department of Veterinary Microbiology and
Immunology, University; of Guelph, Ontario, Canada.
SO Am J Vet Res. 1995 Jul;56(7):841-8. Unique Identifier : AIDSLINE
MED/96039854
AB The core protein and the transmembrane protein, encoded for the
structural genes gag and env, respectively, of caprine
arthritis-encephalitis virus were amplified by use of polymerase chain
reaction, cloned into a pGEX-2T vector, and expressed in Escherichia
coli as fusion proteins with the glutathione S-transferase at their
C-terminus. The recombinant proteins were purified and evaluated by use
of an ELISA. Sera from 269 goats were tested, and the results were
compared with those obtained by use of immunoblot analysis. When results
from both recombinant ELISA (r-ELISA) were compared, it appeared that
the transmembrane glycoprotein was more immunoreactive than the core
protein, because it was recognized by a higher percentage of sera from
infected goats. When results of the 2 ELISA (p28 r-ELISA and p40
r-ELISA) were combined in parallel, they were comparable to those of the
immunoblot test, with sensitivity of 100% and specificity of 98.3%. It
was also found that use of both r-ELISA makes it possible to compare the
variable immunoreactivity against gag and env viral antigens, which may
be correlated with the disease state. The r-ELISA, using core and
transmembrane proteins, appears to be highly sensitive and specific for
detection of antibodies against caprine arthritis-encephalitis virus.
DE Animal Arthritis-Encephalitis Virus, Caprine/GENETICS/*ISOLATION &
PURIF Base Sequence Cloning, Molecular Comparative Study DNA Primers
Electrophoresis, Polyacrylamide Gel Enzyme-Linked Immunosorbent
Assay/METHODS Escherichia coli False Positive Reactions Gene
Products, env/ANALYSIS/*BIOSYNTHESIS/ISOLATION & PURIF Gene Products,
gag/ANALYSIS/*BIOSYNTHESIS/ISOLATION & PURIF *Goat Diseases Goats
Lentivirus Infections/DIAGNOSIS/*VETERINARY Molecular Sequence Data
Polymerase Chain Reaction Recombinant Fusion
Proteins/ANALYSIS/BIOSYNTHESIS/ISOLATION & PURIF Reproducibility of
Results Sensitivity and Specificity Support, Non-U.S. Gov't JOURNAL
ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).