Peste Des Petits Ruminants Virus (PPRV) Ab ELISA Kit for Ovine 96
Wells/Kit
1. Usage
It is used to detect PPRV antibody in serum and plasma of sheep and
goat qualitatively, evaluate the immune status of Peste des petits
ruminants vaccine and assisted serological diagnosis of infected
animals.
2. Principle
This kit use competition ELISA method, PPRV antigen is pre-coated
on enzyme micro-well strips. When testing, add diluted serum sample
and Monoclonal Enzyme conjugate, after incubation, if there is PPR
virus specific antibody, it will bind to the PPRV antigen on the
coating plate and prevent the enzyme-labeled monoclonal antibody
from binding to the antigen on the plate; conversely, if the sample
does not contain PPRV-specific antibody, it will not bind to the
coating plate. After washing to remove the unbound antibody and
other components, add substrate to the microwells to form a blue
product through enzymatic catalysis. After adding stop solution to
terminate the reaction, use a microplate reader at 450nm/630nm
double wavelength to measure the absorbance A value in the reaction
well.
3. The kit components
| 1 | PPRV antigen coated microplate | 96T X 2 |
| 2 | Enzyme conjugate | 12ml | yellow lid |
| 3 | Sample dilution | 25ml | transparent lid |
| 4 | PPRV-IgG Negative control serum | 1ml | green lid |
| 5 | PPRV-IgG Positive control serum | 1ml | red lid |
| 6 | Substrate | 12ml X2 | orange lid |
| 7 | Stop solution | 12ml | blue lid |
| 8 | 10×concentrated washing buffer | 50ml | white lid |
| 9 | Adhesive Foil | 2 pieces | |
| 10 | Instruction | 1 piece | |
4. Material required not provided
1) Micropipette: 0.5-10ul, 10ul-100ul, 100ul-1000ul.
2) Disposable pipette tips.
3) Graduate: 500ml.
4) Microplate Reader: 96 wells with 450/630nm wavelength.
5) Distilled water or deionized water.
6) Microplate Washer
5. Sample preparation
Take animal whole blood, get serum by using regular method, the
serum should bright and no hemolysis.
6. Washing buffer preparation
Return 10X Concentrated washing buffer into room temperature before
use, If there is salt crystals, put it in at 37℃water bath for
5~10min to dissolve it, then dilute it with deionized water at 10
times (for example, to prepare 200ml washing buffer: 180ml of
deionized water + 20ml of 10X concentrated washing buffer, mix it
evenly). The diluted washing buffer can be stored at 4℃for about a
week.
7. Notes
1) Return all reagents into room temperature before use, put all
reagents at room temperature for at least 30 minutes. Shake it
evenly before use, and store back to 2-8℃after usage.
2) Do not mix use reagents from different kits and different lot
no., prevent the reagents been polluted when using.
3) Substrate and stop solution may have irritation to skin and
eyes, be careful to use.
4) Do not expose Substrate to strong light and avoid contact with
the oxidant.
5) PPRV-Ag coated plates should be sealed and moisture-proof. Put
back unused MicroWell plate into dry foil bag and sealed at 2-8 ℃.
6) All wastes should be treated well to avoid pollution before
discarding.
7) Strict compliance with the operating instructions can get the
best results. Pipetting operation, timing, and washing of the whole
process must be precise.
8. Test procedure
1) Take the pre-coated microplate (according to the number of
samples, it can be disassembled and used separately), set 2 wells
for positive control, 2 wells for negative control , add 50µl
positive control serum or negative control serum to it’s well
accordingly; others are sample well, firstly add 40µl sample
dilution, then add 10µl sample into each well. At last, add 50µl
Enzyme conjugate to each well, shake gently to mix it evenly, cover
the plate with adhesive foil,incubate at 37 ℃ for30 minutes;
2) Open the adhesive foil, discard the liquid of the well, add
diluted washing buffer to each well, 300ul/well, be static for 30s,
then discard the liquid, repeat the above step for 5 times, at last
flap to dry with the absorbent paper;
3) Add substrate solution,100ul/well, mix it evenly then cover it
with Adhesive Foil,incubate at 37 ℃in darkfor15 minutes;
4) Add stop solution 50ul/well to stop the reaction, measure the
result in 10 minutes.
9. Results judgement
Read the OD value with ELISA Reader at 450nm (630nm as reference).
For the assay to be valid:
OD value of Negative control (N)≥1.0,
OD value of positive control (P) ≤0.3;
Calculate method:
Sample OD value/average OD value of Negative control (N)= S/N value
Results interpretation
S/N value≥0.5: Negative
S/N value<0.5: Positive
10. Storage and expire date
Store at 2~8℃ in dark, no frozen, expiry date: 12 months.
Q1: How long will it take for you to ship?
A1: Usually ships within 7 working days.
Q2: Do you support OEM/ODM?
A2: can be supported. We can customize according to your specific
needs and specific quantities.
Q3: How is your factory doing in terms of quality control?
A3: We have ISO9001 certification and ISO13485 certification, so
far all the production process, we have standard rules, we comply
with the relevant behavior and laws of the government.
Q4: Is after-sales service guaranteed?
A4: We provide professional online technical after-sales service.
If the product fails during the experiment, we can provide
one-to-one guidance through telephone, video and other forms.
Q5: What is your minimum order quantity?
A5: 1Kit.
Q6: What is the shipping method?
A6: It can be shipped by express (FEDEX, UPS, DHL, EMS, etc.) or by
air and ground.Please confirm with us before placing an order.
