[Animal Modeling]-Immune magnetic bead separation technology combined with flow cytometry to detect Pig-a gene mutations in rats caused by ENU

  Purpose: Combine immunomagnetic bead separation technology and flow cytometry to concentrate and detect rat peripheral blood and Pig? The mutant red blood cells. a Optimize the method of testing gene mutations.

  Methods: Using 20 SD rats, 40 mg and 80 mg/(kg·bw) N-ethyl-N-nitrosourea (N-ethyl-N-nitroso Urea, ENU). Have you collected the peripheral blood of rats from 7 to 14 days and 28 days, concentrate RETCD59- and RBCCD59- on the immunomagnetic separation column, and analyze the peripheral blood before and after the immunomagnetic separation column by flow cytometry The red blood cells were counted?

  Result: Compared with the control group, the incidence of RETCD59- and RBCCD59- in each ENU dose group increased significantly (P\u003c0.05). After using immunomagnetic separation technology, Pig? The sample will be completed in 3 minutes. For gene mutation detection, does the number of RETCD59- or RBCCD59- cells analyzed reach 2x104 or 9x104, respectively?

  Conclusion: This study established and optimized the combination of immunomagnetic bead separation and flow cytometry. Is the pig in the mouse? Can genetic mutation testing methods achieve high-throughput testing and improve testing accuracy and efficiency?