EBV-VCA IgA Ab ELISA Kit Catalog No.: BE501A Enzyme immunoassay for the detection of IgA antibody to the viral
capsid antigen (VCA) of Epstein-Barr Virus (EBV) .
1. PRINCIPLE OF THE ASSAY The EBV-VCA IgA Ab ELISA Kit utilizes a detection system where
microplate wells are coated with recombinant antigen corresponding
to a highly antigenic segment EBV-VCA. Serum or plasma specimens,
controls are added to the wells. During incubation, IgA antibodies
specific for EBV-VCA present in the specimen will bind to the
recombinant antigen fixed onto the microplate wells. The wells are
washed to remove unbound materials, and anti-human IgA peroxidase
conjugate will bind to the antigen-antibody complex and excess
unbound enzyme conjugates are again removed by washing. The enzyme
substrate, tetramethylbenzidine (TMB), is added upon incubation the
substrate will be hydrolyzed by the bound enzyme and a blue or
blue-green colour develops in wells containing EBV-VCA IgA
antibodies. The enzyme reaction is stopped by the addition of
sulphuric acid. The intensity of colour developed is read
spectrophotometrically at 450nm (450 nm/630 nm) and is proportional
to the amount of antibodies present in the specimen. REAGENTS
2. Materials provided with the kits:
Item
Description
96T
480T
1
Microtiter Well
1
5
2
Negative Control
1ml
5ml
3
Positive Control
1ml
5ml
4
Sample Dilution
12ml
60ml
5
Enzyme Conjugate
12ml
60ml
6
Wash Buffer Concentrate (20x)
30ml
150ml
7
Substrate Solution A
6ml
30ml
8
Substrate Solution B
6ml
30ml
9
Stop Solution
6ml
30ml
10
Plastic Bag
1
5
11
Seal Paper
3
15
12
Manual
1 copy
5 copy
3. ASSAY PROCEDURE
Bring ELISA Kit (all reagents), and Specimens to room temperature
before use (approximately 30 minutes).
Dilute concentrated wash buffer 1:19 with ddH2O. For example, 5 ml
of wash buffer concentrate should be diluted to a total volume of
100 mL with deionized or distilled water.
For each test, set one blank well as background control, two
positive and three negative controls. Dispense 100ml positive
control and negative control duplicate into individual wells.
Neither samples nor HRP-Conjugate should be added into the Blank
well.
Dispense 100ml of Sample Dilution into individual test wells except
the controls.
Add 10ml of each test sample into the test wells; vortex to mix.
Incubate for 30 minutes at 37°C
Wash each well 5 times by filling each well with diluted wash
buffer, then inverting the plate vigorously to get all water out
and blocking the rim of wells on absorbent paper for a few seconds.
Add 100ml of Enzyme Conjugate to each well. Mix it gently by
swirling the microtiter plate on flat bench for 1 minutes. Do not
add Enzyme Conjugate to the blank well.
Incubate for 20 minutes at 37°C
Wash the plate 5 times as step 7.
Add one drop (50ml) of Substrate Solution A (HRP-substrate) to each
well, then add one drop (50ml) of Substrate Solution B (TMB) to
each well. Mix gently and incubate at 37°C for 30 minutes. .
Add one drop (50ml) of Stop Solution to each well to stop the color
reaction. Read the OD value at 450 nm/630 nm with dual filter plate
reader. It is option to read the OD value at 450 nm with single
filter plate reader. (using the OD value of the blank well to
correct all the OD reading from all wells)
Company Profile
Biovantion Inc is independent from international department of
Bioneovan Co., Ltd., which established in 2 0 0 5, is an in vitro
diagnostic reagents manufacturer engaged in the research,
development, production, and focus on clinical testing for
infectious diseases, such as Hepatitis virus, Noro virus, Parvo
virus test kits, our products include rapid test cassette and Elisa
kits.
All our products have been registered in National Medical Products
Administration of China, in year 2020 we get Certificate of ISO
13485:2016 – QUALITY MANAGEMENT SYSTEMS FOR MEDICAL DEVICES from
SGS. More importantly, we have awarded the registration certificate
from National Medical Products Administration for Diagnostic Kit
for IgM Antibody to COVID-19 and IgG antibody to COVID-19 during
the outbreak in China, and also we developed COVID-19 Antigen test
; Neutralizing Antibody Test, these products selling well all
overthe world