Indirect ELISA method for detecting haemophilus parasuis antibody by recombining Neu protein and kit of method
A technology for detection of Haemophilus suis and protein, which is applied in the field of indirect ELISA method and its kit, can solve the problems of lack of cross-protection, risk of spreading poison, high detection cost, etc., to avoid a large number of false negatives and false positives, and avoid spreading poison Danger, the effect of reducing the cost of detection
Examples
Embodiment 1
[0044] The concrete steps of the indirect ELISA method of its recombinant Neu protein detection Haemophilus parasuis antibody are as follows:
[0045] S1: Dilute the prepared antigen to 100ng / 100μl with 0.05 mol / L pH 9.6 carbonate buffer, coat the microplate plate, 100uL per well, lightly shake the microplate plate to spread the antigen evenly on the bottom of the well, 37 ℃ for 2 h, transfer to 4 ℃ for overnight adsorption;
[0046] S2: The next day, wash with PBST washing solution 3 to 5 times, each time for 5 minutes, and pat dry;
[0047] S3: Then block with 0.15% BSA, 200uL per well, incubate at 37°C for 1h, then wash the plate with washing solution for 3 to 5 times;
[0048] S4: Add the serum to be tested diluted with PBS, 100uL per well, set up negative and positive control wells at the same time, act at 37°C for 1 hour, wash the plate as above;
[0049] S5: Add diluted SPA-HRP, act at 23 °C for 1 hour, wash the plate as above;
[0050] S6: Add freshly prepared OPD-H...
Embodiment 2
[0054] The concrete steps of the indirect ELISA method of its recombinant Neu protein detection Haemophilus parasuis antibody are as follows:
[0055] S1: Dilute the prepared antigen to 25ng / 100μl with 0.05 mol / L pH 9.6 carbonate buffer, coat the microtiter plate, 100uL per well, lightly shake the microtiter plate to spread the antigen evenly on the bottom of the well, 37 ℃ for 2 h, transfer to 4 ℃ for overnight adsorption;
[0056] S2: The next day, wash with PBST washing solution 3 to 5 times, each time for 5 minutes, and pat dry;
[0057] S3: Then block with 0.15% BSA, 200uL per well, incubate at 37°C for 1h, then wash the plate with washing solution for 3 to 5 times;
[0058] S4: Add sample after sealing, add 100uL of serum to be tested diluted with PBS to each well, set up negative and positive control wells at the same time, act at 37 °C for 1 hour, wash the plate as above;
[0059] S5: Add diluted SPA-HRP, act at 23 °C for 1 hour, wash the plate as above;
[0060] S6...
Embodiment 3
[0064] The concrete steps of the indirect ELISA method of its recombinant Neu protein detection Haemophilus parasuis antibody are as follows:
[0065] S1: Dilute the prepared antigen to 50ng / 100μl with 0.05 mol / L pH 9.6 carbonate buffer, coat the microplate plate, 100uL per well, lightly shake the microplate plate to spread the antigen evenly on the bottom of the well, 37 ℃ for 2 h, transfer to 4 ℃ for overnight adsorption;
[0066] S2: The next day, wash with PBST washing solution 3 to 5 times, each time for 5 minutes, and pat dry;
[0067] S3: Then block with 0.15% BSA, 200uL per well, incubate at 37°C for 1h, then wash the plate with washing solution for 3 to 5 times;
[0068] S4: Add sample after sealing, add 100uL of serum to be tested diluted with PBS to each well, set up negative and positive control wells at the same time, act at 37 °C for 1 hour, wash the plate as above;
[0069] S5: Add diluted SPA-HRP, act at 23 °C for 1 hour, wash the plate as above;
[0070] S6...
PUM
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Description
Claims
Application Information
- IPC
- G01N33/569; C12N9/24
- CPC
- G01N33/56911; C12N9/2402; C12Y302/01018; G01N2333/924
- Inventors
- 刘俊琦