Shenzhen Wensidun Technology Co., Ltd. |
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Brucellosis (Brucella) Antibody ELISA Test Kit for Bovine,Sheep,Pig and Dog 96Wells/Kit ISO9001
1. Diagnostic ELISA Kit Introduction
The Brucellosis antibody ELISA kit is used to test Brucellosis antibody in serum of bovine, sheep, pig and dog etc.
This kit use competitive ELISA method, Brucellosis antigen is pre-coated on enzyme micro-well strips. When testing, add diluted serum sample, after incubation, if there is Brucellosis specific antibody, it will combine with the pre-coated antigen, discard the uncombined antibody and other components with washing; then add enzyme labled anti-Brucellosis virus monoclonal antibody, antibody in sample block the combination of monoclonal antibody and pre-coated antigen; discard the uncombined enzyme conjugate with washing; Add TMB substrate in micro-wells, the blue signal by Enzyme catalysis is in inverse proportion of antibody content in sample, use ELISA reader at 450nm wavelenth to measure the absorbance A value in reaction wells after adding stop solution to stop the reaction.
2. Reagents and contents
Code | Item | Spec. | Code | Item | Spec. |
1 | Brucellosis-Ag Coated plates | 96 wells | 6 | Stop solution | 6 ml |
2 | Enzyme Conjugate | 6 ml | 7 | Negative control | 150 ul |
3 | 10X Concentrated washing buffer | 100 ml | 8 | Positive control | 50 ul |
4 | Substrate | 11 ml | 9 | Adhesive Foil | 2 pieces |
5 | Sample diluent | 20 ml | 10 | Instruction sheet | 1 piece |
3. Material required not provided
1) Micropipette: 10ul-100ul, 100ul-1000ul.
2) Disposable pipette tips.
3) Graduate: 500ml.
4) Microplate Reader: 96 wells.
5) Distilled water or deionized water.
6) Microplate Washer
4. Sample preparation
Take animal whole blood, get serum by using regular method, the serum should bright and no hemolysis
5. Washing buffer preparation
Return 10X Concentrated washing buffer into room temperature before use, if there is salt crystals, shake to make it dissovled, then dilute it at 10 times with distilled water or deionized water. The diluted washing buffer can store at 4℃ for about 1 week.
6. Notes
1) Return all reagents into room temperature before use, 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) Brucellosis-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) Brucellosis-Ag coated plates is disposable, do not repeat use.
7. Test procedure
1) For every test, set 1 well for positive control and 2 wells for negative control;
2) Add sample diluent into each well, 40ul/well;
3) Add serum, negative control and positive control to its corresponding wells, 10ul/well. (Do not mix use the tips);
4) Adding Enzyme Conjugate,50ul/well, shake gentle to mix it evenly, cover it with Adhesive Foil, incubate at 37 ℃ for 60 minutes;
5) Open the adhesive foil, discard the liquid of the well, add diluted washing buffer to each well, 250ul/well, discard the liquid, repeat the above step for 4-6 times, at last flap to dry with the absorbent paper;
6) Add substrate, 100ul/well, mix it evenly then cover it with Adhesive Foil, incubate at 37 ℃ in dark for 15 minutes;
7) Add stop solution, 50ul/well to stop the reaction, measure the result in 10 minutes.
8. Results judgement
Set zero at blank control well, read the OD value at 450nm (630nm as reference).
For the assay to be valid:
OD value of negative control(N) > 0.5, meanwhile positive value (P) blocking rate > 80%
Calculate method:
PI(blocking rate)= {1- (Sample OD value/ Negative control OD average value)}x 100%
Results interpretation
PI(blocking rate)> 70%: Positive
PI(blocking rate)≤ 70%: Negative
9. Storage and expire date
Store at 2~8℃ in dark, no frozen, expiry date: 12 months.
Clinical features
1. Pregnant cows often have miscarriage, stillbirth or weak fetuses
at 6-8 months of gestation. After miscarriage, they often discharge
dirty gray or brown lochia. Some cases have retained placenta,
endometritis, and dirty brownish red exudate injected from vagina.
Damage to the breast may cause a decline in the amount of milk
produced. In the severe cases, the milk may be completely
deteriorated, the breast may become hardened, or even lose the
ability to produce milk; It is also accompanied by arthritis. Bulls
are mainly characterized by testicular and paraorchitis.
2. Abortion of sheep and sheep can cause arthritis and synovial
bursitis. Rams develop orchitis.
3. The main causes of pigs are abortion, endometritis and
infertility, stillbirth and weak offspring. Acute or chronic
orchitis or epididymitis occurs in boars.
Characteristics of pathological changes
It can be seen that fetal septicemia changes, inflammatory
reactions (exudation, necrosis, suppuration or caseification) of
tissues and organs (uterus, breast, placenta, testis and
epididymis) and cell proliferation form granuloma (nodules are
composed of epithelioid cells and macrophages) to scarring.
Immunoserology
There are many serological techniques that can be used for the
diagnosis of brucella, among which the most widely used serum
agglutination test and complement binding test can also be used
because of their significance in differential diagnosis. Other
serum methods such as tiger red plate agglutination test and
antiglobulin test can be used as auxiliary diagnosis.
Allergy
Generally, skin allergy occurs about 20-25 days after brucellosis
infection. This method is not suitable for early diagnosis, but can
be used for mass quarantine.
Microbiology
Samples can be taken from the uterus, vaginal secretions, blood,
viscera, stomach contents, liver, spleen, lymph nodes, and blood of
the aborted female for microbiological examination (staining
microscopy, isolation and culture, animal inoculation).
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.