动物造模,疫缺陷病毒 z-bio 2022-06-01 271

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

　　Methods: The recombinant expression plasmid of SIV SIV30 gene was constructed using the prokaryotic expression vector pGEX-4T-1, and expressed in competent cells BL21. The recombinant protein was purified and used as a diagnostic antigen, and a standardized detection program of immune comb was established, which was applied in clinical practice. detection.

　　Results: The optimal amount of antigen coating was 0.02 mg/mL; the prepared IC could specifically detect the corresponding SIV positive serum without cross-reacting with other virus serum, indicating that the detection method has strong specificity and sensitivity. The analysis results showed that the SIV30 protein could sensitively detect SIV positive serum diluted 1:400 times; the stability and repeatability test results showed that the same sample was repeatedly tested three times, and the coefficient of variation (CV) was less than 10%; Methods 10 suspicious monkey serum samples were tested, the consistency rate between IC and ELISA test results was 100.0%, Kappa = 1:000.

　　Conclusion: The SIV30 protein was expressed in prokaryotic cells, and IC was prepared and applied in clinical detection. The detection method has high sensitivity, strong specificity, good repeatability and good practicability.