Canine (dog) hepatitis E virus IgG,HEV-IgG ELISA Kit, 96T
Purpose
This kit is used for the
determination the existence of Canine Hepatitis E virus IgG (HEV-IgG) in canine
serum,
plasma,
cell culture supernatants,
and other biological fluids.
Principle of the assay
The kit
assays canine HEV-IgG
in the samples by the method of purified HEV antigen pre-coated enzyme-labeled
SPA ELISA. The solid-phase antigen is prepared by purified canine HEV antigen
which is pre-coated onto a microplate. After loading of samples, the HEV-IgG in
the controls or samples bind with the coated antigen. Then the
Staphylococcal protein A (SPA)-enzyme immunoglobulin conjugates has been added
into wells. After washing to remove any unbound reagent, the
tetramethylbenzidine (TMB) substrate is added. TMB substrate produces blue color
when catalyzing by the HRP enzyme. After reaching the desired color intensity,
the reaction is terminated by addition of an acidic Stop solution which changes
the solution color from blue to yellow. Determining the absorbance (OD Value) at
wave length 450 nm by the microwell reader, and the existence of HEV-IgG
can be calculated according to OD from positive or negative controls.
Calculation
1. Average the
duplicate readings for each control and sample and subtract the
average of blank well (1 x sample buffer only).
2. Determine
Cutoff Value (COV): COV= ODnegative + 0.22.
3. ODpositive
or ODnegative is the OD450 from positive or negative
control.
4. If ODsample
≥ COV, the testing sample is HEV-IgG positive; if ODsample < COV, the
testing sample is HEV-IgG negative.
5. The normal
range of ODnegative should be less than 0.1, and ODpositive
should be more than 1.0, otherwise the experiment may have big errors and
maybe need repeating.
6. If the ODsample
is more than 2.5 or less than 0.1, it is out of the
optimal
test range. Dilution or concentration of sample may be necessary for accuracy.
Kit Performance
1.
Sample from healthy
dogs should be negative, e.g.
ODsample <
0.3.
2.
Specificity:
simultaneous detection of recombinant or natural canine
HEV-IgG
without cross-reaction with other cytokines.
3.
Repeatability: both
inter-assay and intra-assay were less than 10 %.
*Lead time of this kit is 1-2 weeks. Shipped by
FedEx air.
*Storage: 2 - 8 C. Shelf Time: 6 months.
*Kit Experimental Time:< 2 hs
>>Trial pricing is available for first time use for certain customers,
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Important note:
Products are intended for laboratory or
manufacture use and are not to be used for diagnosis or any other purpose, which
include but are not limited to, unauthorized commercial uses, ex vivo or in
vivo therapeutic uses. Not for Human or Animal Consumption.