TITLE:
Enzyme-Linked Immunosorbent Assays Using the Recombinant gp51 and p24 of Bovine Leukemia Virus for Immunodetection of the Disease
AUTHORS:
Alejandra Larsen, Santiago Corva, Javier Panei, Christoph Geisler, Eduardo Mortola
KEYWORDS:
Bovine Leukemia Virus, Indirect Dual ELISA, Recombinant Antigens
JOURNAL NAME:
Open Journal of Animal Sciences,
Vol.7 No.3,
July
6,
2017
ABSTRACT: Enzyme-linked immunosorbent assay (ELISA) is often
used to test bovine leukemia virus (BLV) infection. However, commercially
available kits test in South America detect only antibodies against the gp51
protein. With the aim to improve the sensitivity of the test, we developed here
a two-step indirect dual ELISA test that included both proteins p24 and gp51,
expressed and produced in E. coli and
baculovirus expression system respectively. Two hundred ten BLV sera, stated as
double positive or double negative by the combination of commercial agar gel
immunodiffusion (AGID) assay and a gp51-ELISA test, were tested with our in
house dual rp24/rgp51 ELISA. Firstly, we checked the purified, optimized and
standardized proteins as antigen by the checkerboard technique, and set up our
in house ELISA test. The concordance
correlation coefficient (CCC) and coefficient of variation (CV) intraplate
repeatability levels were within the values established by the international
standards. The statistical analysis demonstrated the value of sera correctly
ranked highest (93.48%), and for 0.3 cutoff, the sensitivity was 95.65% and the
specificity 91.30%. In conclusion, the rp24/rgp51 ELISA developed and
standardized here demonstrated to have good analytical characteristics to be
considered for screening of BLV.