动物造模 z-bio 2022-07-13 290

　　OBJECTIVE: To enrich and detect mutant red blood cells in peripheral blood of rats by combining immunomagnetic bead separation technology and flow cytometry, and to optimize the pig-a gene mutation test method in peripheral blood of rats.

　　Methods: SD rats were administered with N-ethyl-N-nitrosourea (ENU) at doses of 20, 40 and 80 mg/(kg·bw) for 3 consecutive days. The peripheral blood of rats was collected on the 7th, 14th and 28th days after exposure, and the RETCD59- and RBCCD59- were enriched by immunomagnetic separation column. Count.

　　Results: Compared with the control group, the incidences of RETCD59- and RBCCD59- in each dose group of ENU were significantly increased (P<0.05). After using the immunomagnetic bead separation technology, the Pig-a gene of a sample could be completed within 3 minutes. For mutation detection, the number of RETCD59- or RBCCD59- cells analyzed was up to 2 × 104 or 9 × 104, respectively.

　　Conclusion: In this study, a combination of immunomagnetic bead separation and flow cytometry was used to establish and optimize the pig-a gene mutation test method in rats, achieve high-throughput detection, and improve the accuracy and efficiency of the test.