Duck viral enteritis virus IgG antibody ELISA test kit instruction manual

Duck viral enteritis ( DVE ), also known as duck plague ( DP ), is an acute, heat, and septic infection of ducks, geese, and various geese-shaped poultry caused by duck viral enteritis virus ( DEV ). Rapid spread, high morbidity and mortality, causing huge economic losses to the poultry industry in China. After immunization prevention of duck virus enteritis, duck virus enteritis inoculated with the vaccine, due to a variety of factors could cause the immune duck does not produce or produce low levels of duck enteritis virus antibodies.

gB protein is a viral envelope protein with adsorption, penetration, fusion cells and intercellular communication. It has good reactogenicity and immunogenicity, and induces high levels of humoral and cellular immune responses. The kit uses an enzyme-linked immunosorbent assay to detect anti- duck viral enteritis virus gB protein antibody IgG in duck serum . It is used to monitor the dynamic level of antibodies after immunization of duck viral enteritis vaccine, to guide the work of immunization prevention, to evaluate the immune status of duck viral enteritis , and also to be used for epidemiological investigation of the disease.

【product content】

[ingredient traits]

[Usage and judgment]

1 , need to bring their own equipment

1.1 micropipette;

1.2   a pipette used with a micropipette;

1.3   a 500 ml measuring cylinder used to dilute the washing solution ;

1.4   Suitable for the detection of 96T microplates;

1.5   a 1.5 ml Ep tube used to dilute the sample ;

1.6   Distilled or deionized water.

2 , sample preparation

The sample to be tested is diluted 100- fold with the sample diluent (recommended: 2 μl of the serum sample is added to the 198 μl sample dilution), and the sample is thoroughly mixed before being added to the coated plate, and the different samples to be tested are diluted to pay attention to the replacement of the tip. Note: 1 blood samples for serum preparation should be free of hemolysis; 2 can not be diluted negative, positive control.

3 , the operation steps   (* Mix gently by shaking or shaking before use, all reagents should be returned to room temperature )

3.1   A1 and A2 in the wells was added 100μl negative control, respectively;

3.2   A3 and A4 in the wells was added 100μl positive control, respectively;

3.3   Take 100 μl of the sample to be tested diluted 1 : 100 into the corresponding well and record it;

3.4   Incubate at 37 °C for 1 h ;

3.5   Discard the liquid from each well into the waste container, add 250μl of washing solution to each well ( 20× concentrated washing solution in the kit should be diluted with distilled water or deionized water before use. If there is crystal, dissolve it before you can dissolve it. Dilute) to wash the plate, repeat 5 times, each time 30s ;

* Prevent the flow of washing liquid between the holes to affect the detection result when adding liquid, and try to throw the washing liquid in the hole as much as possible;

3.6   Add 100 μl of enzyme-labeled conjugate per well ;

3.7   Incubate at 37 °C for 1 h ; repeat step 3.5 ;

3.8   Add 100 μl of substrate solution to each well ;

3.9   Incubate at 37 °C for 10 min (protected from light) ;

3.10   Add 100 μl of stop solution to each well ;

3.11   The absorbance value of each well, i.e., the OD ₄₅₀ value , was measured immediately at a wavelength of 450 nm .

4 , test effectiveness judgment

4.1 Negative control: Under normal circumstances, the negative control well OD ₄₅₀ value ≤ 0.2 ;

4.2 Positive control: Under normal circumstances, the positive control well OD ₄₅₀ value ≥ 0.4 ;

4.3 threshold (CO) calculated: threshold value = mean of negative controls + 0.2; negative control OD ₄₅₀ value greater than 0.2 should be discarded, if all negative control OD ₄₅₀ value greater than 0.2 is to be repeated experiment; negative control is less than 0.05 in 0.05 calculation.

4.4 Determination of results:   Detecting sample OD ₄₅₀ value ≥ critical value, determining that the test sample is positive;   The sample OD ₄₅₀ value <detection value was detected, and it was judged that the test sample was negative.

【Precautions】

( 1 ) When the number of serum samples to be tested is too large, all the serum to be tested should be diluted with a serum dilution plate, and then the serum samples are uniformly added to the microplate to make the reaction time uniform.

( 2 ) Do not smoke, drink or eat during the use of the kit.

( 3 ) The substrate liquid and the stop solution are irritating to the skin, and attention is paid to protection.

( 4 ) Do not expose the substrate to glare and any oxidizing agents; use a clean glass and tanning container to treat the substrate.

( 5 ) All reagents should be stored at 2 ~ 8 °C ; return to room temperature before use, put back 2 ~ 8 °C after use .

( 6 ) Take care to prevent contamination of the kit components.

( 7 ) Do not use reagents that exceed the expiration date. Do not mix the components of different batches of kits.

( 8 ) Pipetting, time and washing during operation must be precise.

[Specifications and packaging] 96T × 2

[Storage and expiration date] 2 ~ 8 °C protected from light, valid for 6 months

For veterinary diagnostic use only

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