Enzyme-linked immunosorbent assay for the detection of HBsAgin serum or plasma
The HBsAg EIA is a solid-phase simultaneous sandwich immunoassay,
which employs monoclonal antibodies and polyclonal antibodies
specific for HBsAg. Microtiter well are coated with monoclonal
antibodies specific for HBsAg. A serum specimen is added to the
antibody coated Microtiter wells together with enzyme conjugated
polyclonal antibodies. HBsAg, if present, will form an
antibody-HBsAg-antibody-enzyme complex. The plate is then washed to
remove unbound material. Finally, a solution of substrate is added
to the wells and incubated. A blue color will develop in proportion
to the amount of HBsAg present in the specimen. The
enzyme-substrate reaction can be stopped and the result is
visualized by naked eye or read by EIA plate reader for absorbance
at the wavelength of 450 nm(450 nm/630 nm) and is proportional to
the amount of antibodies present in the specimen..
Materials provided with the kits:
Item
Description
96T
480T
1
Coating Plate
1
5
2
Negative Control
0.5ml
2.5ml
3
Positive Control
0.5ml
2.5ml
4
Enzyme Conjugate
6ml
30ml
5
Wash Buffer Concentrate (20x)
20ml
100ml
6
Substrate Solution A
6ml
30ml
7
Substrate Solution B
6ml
30ml
8
Stop Solution
6ml
30ml
9
Plastic Bag
1
5
10
Seal Paper
3
15
11
Manual
1 copy
5 copy
ASSAY PROCEDURE:
It is strongly advised to analyze each specimen and controls in
duplicate. All the reagents should equilibrate to room temperature
before use.
Dispense one drop (50μl) of Positive Control as well as Negative
Control in duplicate into respective wells. Set one blank well as
background control, and 50ul of serum or plasma samples into
respective test wells.
Add one drop (50μl) 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.
Place the microtiter plates into a humidified box, and incubate at
37°C for 30 minutes.
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 one drop (50μl) of Substrate Solution A (HRP-substrate) to each
well, then add one drop (50μl) of Substrate Solution B (TMB) to
each well. Mix gently and incubate at 37°C for 15 minutes.
Add 1 drop (50μl) 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