This Document describes in d

1. ubio
quickVET
RAPID
Pregnancy
Test
for
Cattle
u bi o B i ot e c h n ol ogy Sy s t e ms Pv t . Lt d.
Ph : +91-484-2532966, +91-9446438678
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Accurate
diagnosis
of
pregnancy
is
of
crucial
importance
in
maintaining
optimal
reproductive
performance
of
cattle
and
high
profitability
of
dairy
farms.
It
is
desirable
for
the
farmer
to
know
as
soon
as
possible
if
a
mated
or
inseminated
cow
is
not
pregnant,
so
that
she
can
be
rebred
with
the
minimum
delay.
It
is
most
beneficial
if
non-­‐pregnancy
can
be
detected
before
the
first
heat
after
insemination.
Conventional
methods
that
allow
early
detection
of
pregnancy
are
not
accurate
enough,
and
reliance
on
these
methods
results
in
a
large
number
of
non-­‐pregnant
cows
assumed
to
be
pregnant
and
some
pregnant
cows
being
inseminated
inappropriately,
which
could
abort
them.
Conventional
pregnancy
detection
methods
that
are
accurate,
on
the
other
hand,
can
be
performed
only
after
at
least
35
days
after
insemination,
leading
to
the
loss
of
at
least
one
opportunity
to
inseminate.
ubio’s
quickVET
pregnancy
detection
test
provides
a
perfect
tool
to
detect
non-­‐pregnancy
in
cattle
accurately
within
21
days.
The
kit
provides
100%
detection
of
non-­‐pregnant
cases
and
allows
insemination
at
the
first
opportunity.
The
average
savings
realised
through
the
use
of
ubio
quickVET
test
is
as
high
as
Rs.1300/-­‐
per
cow
per
lactation
period.
Moreover,
the
test
can
be
easily
performed
within
20
minutes
at
cow-­‐
side
without
any
instrumentation,
which
makes
the
tool
attractive
even
for
direct
use
by
dairy
farmers.
The
test
employs
the
well-­‐tested
progesterone
assay
principle
to
detect
pregnancy.
It
measures
progesterone
levels
in
milk
or
serum
(for
heifers)
using
a
Rapid
ELISA
technique.
COMPARISON
OF
PREGNANCY
DETECTION
TECHNIQUES
Test
Stage
Principle
Advantages
Disadvantages
Accuracy
Non-­‐return
to
3
wks
Recording
dates
of
-­‐
Early
-­‐
Relies
on
good
About
service
service
and
detection,
oestrus
detection
50%
observation
of
without
missing
symptoms
of
heat
first
heat
after
-­‐
Large
variation
insemination
within
a
population
-­‐
Insemination
at
‘false
oestrus’
can
lead
to
abortion
Early
4
wks
+
Detects
pregnancy-­‐ -­‐
Relatively
-­‐
Needs
blood
About
Conception
dependent
protein
early
detection
-­‐
May
persist
after
90%
Factor
complex
using
ELISA
loss
of
pregnancy
Ultrasound
4
wks
+
Visualization
of
-­‐
Relatively
-­‐
High
capital
cost
About
Scan
images
of
the
early
detection
-­‐
Need
good
100%
conceptus
using
-­‐
Accurate
technical
expertise
realtime
ultrasound
Palpation
5-­‐6
Feeling
the
presence
-­‐
Accurate
-­‐
Possible
damage
About
wks+
of
a
fetus
swelling
in
to
cow
or
conceptus
100%
one
of
the
uterine
-­‐
Need
good
skill
horns
by
inserting
an
and
technical
arm
into
the
rectum
expertise
of
the
cow.
Oestrone
15
Detects
oestrone
-­‐
Very
accurate
-­‐
Too
delayed
to
be
100%
Sulphate
wks+
sulphate
in
milk.
useful
in
most
cases
Concentration
is
proportional
to
oestrogens
produced
by
conceptus.
Conventional
3
wks
Progesterone
-­‐
Accurate
-­‐
High
capital
cost
100%
for

2. Milk
concentration
at
19-­‐ -­‐
Early
-­‐
Requires
blood
in
non-­‐
Progesterone
23
day
interval
from
detection
heifers
pregnancy
plate
ELISA
last
heat
is
much
-­‐
Non-­‐invasive
-­‐
Test
to
be
85%+
for
higher
in
pregnant
performed
on
pregnancy
cows
than
non-­‐ specified
days
pregnant
ones
ubio
3
wks
Progesterone
-­‐
Accurate
-­‐
Requires
blood
in
100%
for
quickVET
concentration
at
19-­‐ -­‐
Early
heifers
non-­‐
Rapid
Milk
23
day
interval
from
detection
-­‐
Test
to
be
pregnancy
Progesteron last
heat
is
much
-­‐
Non-­‐invasive
performed
on
85%+
for
e
Assay
higher
in
pregnant
-­‐
No
specified
days
pregnancy
cows
than
non-­‐ instrumentation
pregnant
ones
-­‐
Cow-­‐side
test
ASSAY
PRINCIPLES
The
ubio
quickVET
pregnancy
test
is
based
on
ELISA-­‐based
progesterone
assay
principle.
The
test
measures
progesterone
concentration
in
milk
or
blood
using
Rapid
ELISA
technique.
A
progesterone-­‐specific
monoclonal
antibody
is
coated
inside
the
test
cassette.
Milk/serum
and
labeled
progesterone
conjugate
are
added
to
the
test
and
control
wells
during
the
test.
The
progesterone
from
the
milk
and
the
labeled
conjugate
will
compete
to
bind
with
the
coated
antibody.
A
chromogen,
that
gives
color,
is
added
to
the
wells
and
it
will
bind
only
to
the
conjugate.
If
the
sample
contains
progesterone
(animal
is
pregnant),
the
test
well
will
be
lighter
than
the
control
well.
If
the
progesterone
level
is
very
low
(non-­‐pregnant
animal),
the
test
well
be
bluer
than
the
control
well.
The
concentration
of
progesterone
in
the
milk
or
In
the
case
of
a
pregnant
cow,
the
progesterone
blood
of
a
non-­‐pregnant
cow
varies
as
shown
concentration
varies
as
given
below.
below,
with
Day
0
corresponding
to
the
occurrence
of
heat.
It
is
assumed
that
insemination
is
appropriately
performed
at
the
time
of
heat.
It
is
obvious
that
there
is
a
significant
difference
between
progesterone
concentration
in
milk/blood
samples
taken
from
pregnant
and
non-­‐pregnant
cows
from
19th
to
23rd
day
after
heat
(and
insemination).
By
checking
the
progesterone
level
during
this
time
frame,
we
can
accurately
identify
non-­‐pregnancy.
This
principle
provides
100%
accurate
results
for
progesterone
levels,
which
translates
to
100%
accuracy
for
non-­‐pregnancy.
However,
there
are
some
corner
cases
where
a
false
positive
may
occur
for
cases
marked
‘pregnant’
by
the
test.
1. If
heat
is
incorrectly
interpreted,
and
insemination
is
done
at
the
wrong
time,
the
test
will
show
the
result
as
‘pregnant’
as
we
are
actually
detecting
the
high
progesterone
level
during
mid-­‐heat,
instead
of
that
caused
by
pregnancy.

3.
2. A
cow
that
is
correctly
judged
pregnant
might
lose
her
conceptus
due
to
various
reasons.
In
this
case,
the
test
is
accurate,
but
the
pregnancy
does
not
result
in
calf-­‐birth.
It
is
suggested
that
the
test
be
repeated
if
any
sign
of
heat
is
seen
after
the
cow
is
judged
pregnant.
3. A
cow
with
luteinized
cystic
follicles
will
produce
progesterone
at
varying
levels,
and
the
test
result
is
not
reliable
in
such
animals
as
it
might
show
positive
or
negative
based
on
the
progesterone
level
on
the
day
of
test.
CLINICAL
APPLICATIONS
1. Ovulation
Test:
ubio
quickVET
rapid
pregnancy
test
for
cattle
can
be
used
for
detecting
ovulation
in
cows
that
do
not
show
regular
symptoms
of
heat.
The
test
should
be
performed
during
the
suspected
heat
period.
A
dark
blue
spot
indicates
heat
and
insemination
can
be
performed
in
6-­‐
18
hours.
A
clear
white
card
indicates
mid-­‐heat
and
the
test
needs
to
be
repeated
only
after
10
days.
A
light
blue
color
indicates
approaching
or
past
heat,
and
a
repeated
test
after
2
days
will
confirm
the
condition.
2. Identification
of
follicular/luteal
cysts:
Ovarian
cysts
are
an
important
infertility
problem
in
dairy
cattle.
It
is
often
difficult
to
differentiate
ovarian
cysts
into
follicular
and
luteal
by
palpation
of
the
ovary
per
rectum.
However,
once
the
presence
of
an
ovarian
cyst
has
been
determined
by

4. palpation,
differentiation
can
be
made
on
the
basis
of
progesterone
concentration.
Cystic
cows
having
a
low
progesterone
concentration
are
considered
to
have
follicular
cysts,
and
those
with
high
concentrations
are
classified
as
having
luteal
cysts.
For
practical
purposes,
differential
diagnosis
of
ovarian
cysts
is
important
in
selection
of
endocrine
therapy.
The
cyst
can
be
differentiated
by
performing
the
test
on
two
samples
taken
7
days
apart
–
a
dark
blue
spot
in
both
tests
indicate
a
follicular
cyst
and
a
clear
white
card
in
both
tests
indicate
a
luteal
cyst.
3. Evaluation
of
endocrine
therapy:
To
evaluate
treatment
of
follicular
cysts
using
GnRH,
ubio
quickVET
rapid
pregnancy
test
can
be
used
to
evaluate
to
progesterone
levels
10
days
after
administration
of
GnRH.
A
light
blue
spot
or
a
clear
white
card
indicates
response
to
treatment.
Similarly,
the
test
can
be
used
to
evaluate
response
to
treatment
of
luteal
cysts
using
Prostaglandin
compounds
as
unobserved
estrus
following
prostaglandin
administration
is
a
common
occurrence.
Response
to
prostaglandin
is
dependent
upon
the
presence
of
a
functional
corpus
luteum.
Therefore,
milk
progesterone
testing
prior
to
and
3
days
after
administration
of
prostaglandin
can
accurately
assess
if
administration
is
justified
(high
concentration
prior
to
administration)
and
if
luteolysis
has
occurred
(low
concentration
on
day
3).
4. Evaluation
of
fitment
for
embryo
transfer:
ubio
quickVET
rapid
pregnancy
test
for
cattle
can
be
used
in
embryo
transfer
programs
to
check
if
there
is
adequate
progesterone
level
in
the
recipient
to
support
the
embryo.
The
recipient
should
show
a
light
blue
test
result
before
an
embryo
is
transferred.
INTERPRETATION
OF
THE
RESULT
The
control
well
is
usually
colorless
or
a
very
pale
blue.
A
test
well
that
is
considerably
bluer
than
the
control
well
indicates
a
non-­‐pregnant
animal
with
low
progesterone
level.
A
test
well
lighter
than
or
of
the
same
shade
as
the
control
well
indicates
a
pregnant
animal
with
higher
progesterone
concentration,
subject
to
caveats.
Slightly
bluer
tinge
in
the
test
well
when
compared
to
the
control
well
indicates
non-­‐pregnant
result
during
approaching
or
past
heat
period
–
in
this
case,
the
test
timing
and
heat
calculation
must
be
checked,
and
the
test
should
be
repeated
if
necessary.
Not
Pregnant
Not
Pregnant
Pregnant
Pregnant

5. TEST
PROCEDURE
1.
Collect
milk
sample
from
cow
in
a
clean
vessel
after
discarding
milk
from
the
first
few
KIT
COMPONENTS
pulls.
For
pregnancy
test,
the
test
should
be
-­‐
One
Aluminium
pouch
containing
done
between
19th
to
23rd
day
after
the
test
cassette.
insemination.
-­‐
Three
droppers
for
transferring
milk
and
reagents.
2.
Keep
the
test
pack
at
26-­‐30ºC
for
1-­‐2
hours
-­‐
One
transparent
vial
marked
‘1’
to
bring
it
to
room
temperature.
Remove
the
containing
reagent
1.
test
cassette
from
Aluminium
pouch
and
keep
-­‐
One
dropper
bottle
marked
‘2’
on
a
flat
surface.
There
are
two
wells
on
the
containing
wash
buffer
(reagent
card,
marked
C
and
T.
2).
3.
Add
three
(3)
drops
of
milk
to
both
the
wells
-­‐
One
opaque
vial
marked
‘3’
(C
and
T)
using
a
dropper
supplied
with
the
containing
reagent
3
test.
Dispose
the
dropper
immediately.
4.
Keep
the
cassette
on
a
flat
surface
for
5
PRECAUTIONS
&
minutes.
Periodically
tap
the
side
of
the
WARNINGS
cassette
for
proper
mixing.
Take
care
not
to
-­‐
Store
in
fridge
at
4-­‐8ºC,
do
not
splash
the
milk
in
the
wells.
freeze
5.
Using
a
fresh
dropper,
Add
three
(3)
drops
-­‐
Do
not
reuse
test
kit.
of
reagent
into
both
the
wells
(C
and
T)
from
-­‐
Use
a
separate
dropper
for
each
the
bottle
marked
Reagent
1.
Let
milk
and
reagent.
reagent
mix
by
gently
tapping
the
cassette.
-­‐
Do
not
mix
components
from
6.
Keep
the
cassette
on
a
flat
surface
for
20
different
kits.
minutes.
Gently
tap
the
cassette
at
4-­‐minute
intervals.
-­‐
Do
not
touch
the
test
wells.
7.
Tip
the
cassette
and
empty
the
wells
into
a
sink.
8.
Fill
the
wells
with
the
wash
buffer
in
the
dropper
bottle
marked
‘Reagent
2’,
tap
gently
and
empty
the
wells
into
the
sink.
Repeat
this
washing
5-­‐6
times.
Shake
off
any
remaining
solution
from
the
wells.
9.
Add
5
drops
of
Reagent
3
in
the
opaque
vial
to
both
the
empty
wells.
Keep
the
test
on
a
flat
surface.
The
result
appears
after
8
minutes,
and
can
be
read
till
15
minutes.