Porcine pseudorabies virus gE and gI antibody chemiluminescence detection kit and application thereof
A technique for chemiluminescence detection and porcine pseudorabies virus, applied in the field of immune detection, can solve the problems of missed detection of infected animals, difficult detection of specific proteins, low sensitivity of ELISA kits, etc., and achieves the effect of good stability
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0026] Example 1 Establishment of chemiluminescence detection kit for porcine pseudorabies virus gE and gI antibodies
[0027] The optimal coating concentration and serum dilution were determined by checkerboard titration. Among them, the gE Nanobody and the gI Nanobody were made with gradients of 2ug / ml, 1ug / ml, 0.5ug / ml, 0.25ug / ml and 0.1ug / ml respectively, and the positive control serum and the negative control serum were made 1:10 respectively. , 1:20, 1:50, 1:100 times dilution. By comprehensively considering the light index and signal-to-noise ratio, the coating conditions were determined as the mixed coating of gE Nanobody protein 0.25ug / ml and gI Nanobody protein 0.25ug / ml. Serum dilution factor is 1:100.
Embodiment 2
[0028] Example 2 Determination of critical value of porcine pseudorabies virus gE and gI antibody chemiluminescence detection kit
[0029]Serum with known background is detected by using the kit of the present invention. The kits were used to detect 150 positive samples and 150 negative samples, calculate the S / P value of each sample, and use SPSS 16.0 software to analyze the S / P value of all serum samples. The ROC curve was constructed using the nonparametric method, and the cut-off point with the largest Youden index was used as the critical point for positive and negative judgments. The sensitivity and specificity of the kit were also determined. In this experiment, the area under the curve was 0.972, the largest Youden index was 0.8600, and the corresponding S / P value was 0.201. Therefore, the critical value of this method is determined as 0.20, and finally the determination standard of this diagnostic kit is determined as: S / P value figure 1 , figure 2 .
Embodiment 3
[0030] Example 3 Sensitivity and specificity detection of porcine pseudorabies virus gE and gI antibody chemiluminescence detection kit
[0031] Dilute the porcine pseudorabies virus positive serum with serum diluent, the gradients are 1:32, 1:64, 1:128, 1:256, 1:512, 1:1024, 1:2048, respectively. Samples are tested directly. Dilute the positive serum samples of swine fever, porcine blue ear, porcine small, porcine ring type 2, porcine Japanese encephalitis B by 1:100 times, and the diluted samples are directly tested, and the positive control serum and negative control serum are set as controls. . The diluted samples were added to the chemiluminescence-coated plate, and reacted at 37°C for 15 min; washed with washing solution for 5 times, and added horseradish peroxidase-labeled goat anti-pig IgG antibody diluted 10,000 times with 5% BSA-PBST. Incubate at 37°C for 15min; then wash with washing solution for 5 times, add luminescent substrate A and luminescent substrate B, re...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More