细胞原位PCR方法用于异性别骨髓移植植入验证

为监测异性别骨髓移植（BMT）治疗某些血液系统疾病和遗传性疾病的植入状态，利用人Y染色体特异性片段扩增引物Y3、Y4，采用非放射性核苷类似物Dig－11－dUTP直接掺入法，对2例急性粒细胞性白细胞进行原位聚合酶链反应（insituPCR）。在显微镜下观察，Y染色体阳性细胞内显示有一深蓝色信号班点，以此识别供者源细胞。1例患者BMT后＋14天供者源细胞占37％，另1例＋28天时供者源细胞达96．53％，均获得植活证据。该方法简便快速，灵敏度高，只需计数大约200个细胞，一般在检查次日即可获得结果。

THE APPLICATION OF CELLULAR IN SITU PCR TO DOCUMENTING MARROW ENGRAFTMENT AFTER BONE MARROW TRANSPLANTATING

To monitor engraftment after allogenic bone marrow transplantation (Allo-BMT) in treatment hematological disease and genetic diseases,cellular in situ polymerase chain reaction (in situ PCR) was carried out on peripheral blood and marrow cells from 2 female patients with acute myelogenous leukemia (AML) after marrow ablative therapy followed by male donor marrow infusion,by use of human Y chromosome specific primer pairs Y3 and Y4,with direct incorporating nonisotopic nucleotide Dig-11-dUTP. A dark-blue signal spot could be visualized under a microscope in the Y-chromosome positive cell, which indicatied the cell of donor origin. In one patient the cells of donor origin was 37% on day-14 posttransplantation and it was 96. 53% on day-28 posttransplantation in another patient. Evidence of engraftment was obtained in both cases. This method is simple,rapid and highly sensitive,only about 200 cells counting from all sides is necessary and the result could be able to obtained on next day after detected by in situ PCR.