A kind of foot-and-mouth disease type A virus SIGA antibody ELISA detection kit and its application
A detection kit and a technology for foot-and-mouth disease are applied in the field of indirect ELISA detection kits for foot-and-mouth disease type A virus sIgA antibody, and achieve the effects of less stress on animals, less labor consumption and easy operation.
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Embodiment 1
[0053] Preparation and assembly of embodiment 1 foot-and-mouth disease type A virus sIgA antibody ELISA detection kit
[0054] (1) Preparation of the kit:
[0055] 1. The design of the screening of the dominant epitope (TB / A) of the foot-and-mouth disease type A virus:
[0056] The structural protein VP1 of foot-and-mouth disease virus is the dominant antigen of the virus. Whether it is the isolated and purified natural VP1 protein or the recombinant expression product, it can induce the body to produce protective neutralizing antibodies, which is type-specific. The full length of the foot-and-mouth disease virus VP1 gene consists of 639 nucleotides, encoding a protein with 213 amino acids, and its main epitopes are concentrated in the 140-160 amino acid and the 200-213 amino acid segment. The present invention uses DNAStar biological software to represent strain A / GDMM / 2013 (GenBank accession number: KF450794.1), A / HuBWH / 2009 (GenBank accession number: JF792355.1), A / GDMM / 20...
Embodiment 2
[0098] Sensitivity, specificity, repeatability experiment of embodiment 2 foot-and-mouth disease type A virus sIgA antibody ELISA detection kit
[0099] 1. Sensitivity experiment:
[0100] Utilize the kit prepared in Example 1 to detect 90 nasal test sub-samples, these samples are the nasal test sub-samples collected after the experimental challenge, before any immunization is carried out, the detection method is the same as that in Example 1 by calculating the positive The detection rate was used to analyze the sensitivity of the method. The results of ELISA showed that 88 of the above samples were positive and 2 were negative, and the positive detection rate was 97.78%. Illustrate that the kit of the present invention has better sensitivity, and the results are shown in Table 4.
[0101] Table 4 ELISA kit sensitivity test
[0102] Detection method Number of samples (parts) Positive number (parts) Negative number (parts) Positive rate (%) indirect EL...
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