VMRD’s highly-sensitive and specific enzyme-
linked, immunosorbent assay (ELISA) kit
detects antibodies to bovine leukemia virus
(BLV) glycoprotein 51 (gp51) in bovine sera.
Sample serum antibodies bind to BLV gp51
molecules attached to the plastic wells of
the microtiter plate. Binding of these serum
antibodies is detected by reaction with
horseradish peroxidase (HRP)-affinity-purified
goat antibodies to bovine immunoglobulins.
Attached HRP-antibodies are detected
by addition of enzyme substrate and
quantitated by subsequent blue color product
development. Strong color development
indicates the presence of antibodies to BLV
gp51 in the sample serum. Very weak or no
color development indicates the absence
of detectable antibodies to BLV gp51 in the
sample serum. VMRD’s Bovine Leukemia Virus
Antibody Test Kit is USDA-approved for export
testing and is available in breakaway stripwell
format. The assay requires that an ELISA plate
reader be used for accurate results.
About Bovine Leukosis
Enzootic Bovine Leukosis (EBL) is an infectious,
non-contagious viral disease of cattle. It is
caused by BLV, an oncogenic delta retrovirus,
which results in proliferation of B lymphocytes.
Infection with BLV may lead to persistent
lymphocytosis and in some adult cattle to the
development of tumors with associated signs.
The spread of disease from the introduction
into a herd may reach enzootic proportions.
Transmission of BLV occurs between animals
primarily by transfer of B lymphocytes in
blood. Trauma, use of common bleeding
needles, and surgical procedures are the main
means of transmission. It is rarely vertically
transmitted. Most BLV infections are inapparent,
with approximately 5% of animals developing
clinical signs. AGID and ELISA tests are used to
identify carrier cattle. Control programs for EBL
include testing and removal of positive animals.
Several European countries which have
instituted eradication programs also require
that imported cattle be free of BLV.
Component 283-2
A. Antigen-Coated Plates 2 plate
B. Positive Control 3.6 ml
C. Negative Control 3.6 ml
D. 100X Antibody-Peroxidase Conjugate 0.3 ml
E. Conjugate Diluting Buffer 30 ml
F. 10X Wash Solution Concentrate 120 ml
G. Substrate Solution 30 ml
H. Stop Solution 30 ml
Test Kit Insert
DIAGNOSTIC TEST KITS | BOVINE
800.222.8673 | www.vmrd.com
CATALOG NO.
284-2
SPECIES SAMPLE
Bovine
SAMPLE
Serum
SENSITIVITY †
98%
SPECIFICITY †
100%
ASSAY TIME
60 minutes
CONFIGURATION
2 stripwell plates
TESTS
182
CATALOG NO.
284-WASH
120mL of
lot-specific 10X
Wash Solution
Concentrate
† See Sensitivity & Specificity
in Perspective on TOC page
BOVINE LEUKOSIS
Bovine Leukemia Virus
Antibody Test Kit
ELISA
Component 284-2
A. Antigen-Coated Plates 2 plate
B. Positive Control 4 ml
C. Negative Control 4 ml
D. 100X Antibody-Peroxidase Conjugate 0.3 ml
E. Conjugate Diluting Buffer 30 ml
F. 10X Wash Solution Concentrate 120 ml
G. Serum Diluting Buffer 60 ml
H. Substrate Solution 30 ml
I. Stop Solution 30 ml
Test Kit Insert
KIT CONTENTS
OVERVIEW OF KIT PROCEDURE
1. Dilute serum samples 1/25 with Serum Diluting
Buffer
2. Transfer 50 µl of each sample and controls into
wells of the Antigen-Coated Plate
3. Incubate 20 minutes at room temperature
4. Wash 3 times with Wash Solution
5. Add 50 µl of Antibody-Peroxidase Conjugate
6. Incubate 20 minutes at room temperature
7. Wash 3 times with Wash Solution
8. Add 50 µl of Substrate Solution
9. Incubate 20 minutes at room temperature
10. Add 50 µl of Stop Solution
11. Read at 620-650 nm
All samples with mean OD values greater than or equal
to the mean OD of the Positive Controls are considered
positive for BLV. All samples with mean OD values less
than the mean of the Positive Controls are negative for
BLV.
For the test to be valid, the mean OD of the Negative
Controls must be less than 0.200. The mean OD of the
Positive Controls must be ≥0.250 and <2.000.
2