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应用四条引物(R_5、R_6、D_3、D_4)通过逆转录聚合酶链反应(RT-PCR)对9例急性早幼粒细胞白血病(AML-M_3)患者异基因骨髓移植(BMT)前后进行了 RAR~α/MYL 融合 mRNA 动态监测。建立的方法敏感度为0.1ng,总 RNA 相当于在10~4～10~5个正常细胞中检出一个 AML-M_3细胞,扩增产物的特异性经 kpnI 酶切法确认。所有9例病例中,3例于 BMT 后由阳性转阴。4例 BMT 后持续阴性表达,此7例患者均获长期无病生存。1例 BMT 后 RAR~α/MYL 融合 mRNA 持续阳性表达,最终出现白血病复发,1例失访。结果说明 RAR~α/MYL 融合 mRNA 是 AML-M_3的特异性标记,其动态检测有助于 AML-M_3患者 BMT 后残留白血病的检测,对 BMT 后白血病复发有预示价值。
Four RARs (RARs) were performed in all 9 acute promyelocytic leukemia (AML-M_3) patients before and after allogeneic bone marrow transplantation (BMT) by using four primers (R_5, R_6, D_3 and D_4) by reverse transcription polymerase chain reaction ~ α / MYL fusion mRNA dynamic monitoring. The sensitivity of the established method was 0.1 ng, and the total RNA was equivalent to detect one AML-M 3 cell in 10 ~ 4 ~ 10 ~ 5 normal cells. The specificity of the amplification product was confirmed by kpnI digestion. Of all 9 cases, 3 were positive to negative after BMT. 4 cases of persistent negative expression after BMT, 7 patients were long-term disease-free survival. After 1 case of BMT, RAR ~ α / MYL fusion mRNA was continuously and positively expressed, finally leukemia recurrence occurred and 1 case was lost to follow-up. The results indicate that RAR α / MYL fusion mRNA is a specific marker of AML-M 3, and its dynamic detection is helpful to the detection of residual leukemia after BMT in AML-M 3 patients, indicating the value of leukemia recurrence after BMT.