Petits Ruminants veterinary diagnostic kits PPRV Ab ELISA Kit For Ovine

Petits Ruminants Veterinary Diagnostic Kits PPRV Ab ELISA Kit For Ovine

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

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.

Clinical symptoms

1. Most acute type

It is common in goats. After an average incubation period of 2 days, the body temperature suddenly rises to 40-41 degrees, or even 42 degrees, with depression, bristling hair, and loss or disappearance of appetite. At the same time, there are tears and serous nasal mucus. Catarrhal inflammation of the mouth, sometimes ulceration of the mucosa, or death before it occurs. However, hyperemia of tooth m is common. First constipation, then massive diarrhea, but no bloody stool. Suffered from sheep failure, temperature drop and sudden death. The course of disease is 5 to 6 days, and the incidence rate is as high as 100%. In case of serious outbreaks, the mortality rate is 100%; In mild cases, the mortality rate does not exceed 50%. The performance of young sheep is severe, with high incidence rate and mortality.

2. Acute type

The incubation period is 3 to 4 days, and the symptoms are the same as the most acute symptoms, but the course of the disease is longer, so the serous and viscous nasal juice turns to mucopurulent, and the nostrils are soon blocked. The mucous membranes of tongue, cheek, upper jaw and pharynx appear ulcer. There is often necrotizing stomatitis. In the mild cases, small red superficial necrotic foci appear, and in the severe cases, the whole oral cavity is affected. The sick sheep are thirsty and do not eat. Persistent jet diarrhea occurs. If you have difficulty breathing and cough, it indicates respiratory infection. Vaginal-vaginitis often occurs in ewes, accompanied by mucopurulent secretion. Pregnant sheep can cause abortion. The course of the disease is 8-10 days. Some died of concurrent disease, some recovered, and some became chronic.

3. Subacute or chronic type

It is common after the acute phase. Early symptoms are the same as above. Nodules and purulent scars around the mouth and nostrils as well as the lower collar are the special symptoms of the late stage of this type, which are easily confused with infectious purulent scars. The course of disease is 10-15 days. Sheep generally show subacute process, and then recover, or do not show symptoms.

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.