摘要
为了探讨实时定量RT PCR(real timeRT PCR)监测儿童AML患者AML1 ETO融合基因的临床价值 ,筛选出AML1 ETO阳性患儿 14例 ,以连续稀释的AML1 ETO质粒作为标准品建立标准曲线 ,应用实时定量RT PCR监测其初治及随访期 5 8份骨髓中AML1 ETO融合基因 ,用SPSS软件进行统计学分析研究其和临床的关系。结果显示 :实时定量RT PCR监测AML1 ETO基因的敏感度可达 10 - 5,重复性好。 12份初治标本的AML1 ETO∶GAPDH比值平均为 0 .6 4 8,该比值与骨髓中幼稚细胞数的比值无相关性。微小残留白血病 (MRD)定量水平检测发现 ,化疗 1个月时 6例患儿AML1 ETO的量无明显下降 (高于 5× 10 - 2 ) ,其中 3例早期复发 ,1例晚期复发 ;另 4例患儿有明显下降 (低于 10 - 2 ) ,均长期存活。化疗 3个月时 5例高于 5× 10 - 3者中 3例复发 ;另 6例低于 5× 10 - 3者中 1例复发。 6个月后持续高于 10 - 3者 3例均复发。MRD定量水平动态检测发现 ,在化疗中MRD持续在高水平 (>5× 10 - 3)者 4例全部临床复发 ,复发时间在 3个月到半年 ;4例持续低于 10 - 3,维持在 10 - 4- 10 - 6 ,一直处于骨髓缓解期 ;1例化疗中从 10 - 5上升到 10 - 3,5个月后临床复发。 3例持续缓解超过 9个月的患儿仍可检测到融合基因 ,约 10 - 5- 10 - 6 。结论 :实时?
This study was aimed to investigate the clinical value of quantification of AML1/ETO fusion transcripts using real-time reverse transcription PCR. Fourteen AML1/ETO positive children out of 52 AML children were selected. A serial dilution of AML1/ETO plasmid was used as a template for the AML1/ETO real-time PCR. AML1/ETO was quantified according to the expression of the GAPDH housekeeping gene at new diagnosis and during/after chemotherapy and transplantation. SPSS statistics was used to analyze the data. The results showed that the ratio of AML1/ETO∶GAPDH expression level at new diagnosis varied in the range 0.219-2.080 (median 0.648) among the patients, without relevance with percentage of blasts. The detection sensitivity was up to the dilution of 1∶105. Six patients showed a slight decline of AML1/ETO (higher than 5×10 -2) at 1 month, three of whom relapsed in the early stage and one later. Five patients had a higher level than 5×10 -3 at 3 months, three of whom relapsed. Four patients with always a higher level than 5×10 -3 all relapsed in early stage. After six months, four out of them with constant low-level expression (10 -4-10 -6) were in continuous complete hematological remission (CCR). In another patient, a rapid rise of AML1/ETO transcripts could be detected at CR stage and he relapsed 5 months later. The AML1/ETO gene expression leveling off by 10 -5-10 -6 could be detected in 3 patients at their complete remission after 9 months. It is concluded that real-time RT-PCR is a suitable approach for quantifying AML1/ETO transcripts in monitoring of AML patients with t(8;21) during/after chemotherapy and provides data of diagnostic relevance.
出处
《中国实验血液学杂志》
CAS
CSCD
2005年第1期76-82,共7页
Journal of Experimental Hematology
