免疫缺陷病毒抗体,动物造模 z-bio 2021-09-24 492

　　Objective: To prepare the SIV30 protein of simian immunodeficiency virus (SIV) by genetic engineering technology as an antigen diagnostic reagent, and establish an immunocomb method (IC) for specifically detecting anti-SIV antibody IgG in experimental monkey serum.

　　Method: Use the prokaryotic expression vector pGEX-4T-1 to construct a recombinant expression plasmid for the SIVSIV30 gene, and express it in the competent cell BL21. Purify the recombinant protein and use it as a diagnostic antigen to establish a standardized detection program for the immune comb. Do you apply it to clinical trials?

　　Result: The best antigen coating amount is 0.02 mg/mL. The prepared IC can specifically detect the corresponding SIV positive sera without cross-reacting with other virus sera, which indicates that the detection method has strong specificity. Sensitivity analysis results show that SIV30 protein can detect SIV positive serum diluted 1:400 with high sensitivity. The stability and reproducibility test results show that the same sample has been repeatedly tested 3 times, and the coefficient of variation (CV) is less than 10%. When using this detection method to detect 10 suspicious monkey serum samples, the matching rate of IC and ELISA results is 100.0%, kappa = 1:000?

　　Conclusion: Prokaryotic expression of SIV30 protein, IC preparation and clinical application test? This test? Is the method highly sensitive? Is it specific? Excellent repeatability and excellent practicality?