微核胞质分裂阻滞细胞(CB-MNT)方法学探讨和比较

The comparison of methods on the cytokinesis block micronucleus cell (CB-MNT) detection

目的微核胞质分裂阻滞细胞(CB-MNT)实验方法学探讨,对CB-MNT和常规法微核(C-MNT)两种检测方法进行比较。方法选取85名从事放射作业人员外周血标本,分别采用CB-MNT法和常规法两种方法培养微核细胞。通过制片染片及镜下检查,识别双核淋巴细胞微核和转化的单核淋巴细胞微核;比较两种方法培养出的淋巴细胞的自发微核细胞率,并进行统计分析。结果采用淋巴细胞微核自发率CB法制片的结果是3.85±8.43;采用常规法制片的结果是1.13±1.60。两种方法得到的自发率差异有统计学意义(t=-6.17,P<0.01)。采用CB法制片获得的双核淋巴细胞核完整膨大,胞质清晰,微核易识别。结论胞质分裂阻滞法优于常规培养法;胞质分裂阻滞法对毒性损伤反映更加敏感;微核细胞更易识别;能减少阅片误差,结果准确可靠。建议作为职业健康体检对遗传损伤监护中实验室中的优先检查方法。

Objective To compare method of the cytokinesis block micronucleus cell（ CB-MNT） and conventional micronucleus（ C-MNT）. Methods The peripheral blood samples from 85 workers exposed to radiation were collected and cultured by two method ： CB-MNT method and conventional method. Through Giemsa staining and the microscopic examination,the micronucleus rate and the mononuclear lymphocyte micronucleus of the dual nuclei lymphocyte were identified.The spontaneous micronucleus cell rate obtained through the two method were compared and analyzed. Results The micronucleus rate of lymphocyte obtained through the CB-MNT method was 3. 85 ± 8. 43,and through conventional method was1. 13 ± 1. 60. There was significant difference between the two（ P〈0. 01）. Continuous plasma membrane and clear cytoplasm were observed in the double nucleus lymphocyte harvested by CB-MNT method. Conclusion Cytokinesis block method is better than the conventional culture method. It is more sensitive to reflect the toxicity. Micronucleus cells can be identified more easily. The result is accurate and reliable. CB-MNT could be recommended as a priority method used for genetic damage monitoring for occupational health examination.