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首页> 外文期刊>Journal of pediatric hematology/oncology: Official journal of the American Society of Pediatric Hematology/Oncology >Real-time quantitative PCR: standardized detection of minimal residual disease in pediatric acute lymphoblastic leukemia. Polymerase chain reaction.
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Real-time quantitative PCR: standardized detection of minimal residual disease in pediatric acute lymphoblastic leukemia. Polymerase chain reaction.

机译：实时定量PCR：标准化检测小儿急性淋巴细胞白血病的最小残留病。聚合酶链反应。

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摘要

PURPOSE: To develop a standardized real-time polymerase chain reaction (PCR) method of quantifying minimal residual disease (MRD) in patients with pre-B acute lymphoblastic leukemia (ALL). PATIENTS AND METHODS: In a series of 24 follow-up bone marrow (BM) samples in 11 patients (14 clonal markers), we performed real-time PCR assays using one consensus and one clone-specific primer for each marker. The markers analyzed included immunoglobulin heavy chain (IgH), T-cell receptor (TCR) and TEL-AML1 rearrangements. RESULTS: We achieved a detection limit of 3.3 x 10(-5) +/- 1.2 x 10(-5) and an accurate quantitation (r = -0.99) limit of 2.0 x 10(-4) +/- 8.8 x 10(-5) blasts. Both inter- and intra-assay reproducibility were exceptional. Additionally, we found comparable results to those of a "gold standard" limiting-dilution PCR assay (r = 0.62). CONCLUSIONS: The IgH, TCR and TEL-AML1 markers can be used as targets by real-time PCR under the same cycling profile, allowing quantitation of MRD in more 95% of patients with pre-B ALL. This standardized, real-time PCR technique should simplify monitoring MRD in clinical trials.
机译：目的：开发一种标准化的实时聚合酶链反应（PCR）方法，以量化B前急性淋巴细胞白血病（ALL）患者的最小残留疾病（MRD）。患者与方法：在11位患者（14个克隆标记）的一系列24个随访骨髓（BM）样本中，我们对每个标记使用一个共有序列和一个克隆特异性引物进行了实时PCR分析。分析的标志物包括免疫球蛋白重链（IgH），T细胞受体（TCR）和TEL-AML1重排。结果：我们实现了3.3 x 10（-5）+/- 1.2 x 10（-5）的检出限和2.0 x 10（-4）+/- 8.8 x 10的准确定量（r = -0.99）限（-5）爆炸。批间和批内重复性均优异。此外，我们发现了与“金标准”有限稀释PCR分析（r = 0.62）可比的结果。结论：IgH，TCR和TEL-AML1标记可以在相同的循环模式下通过实时PCR用作靶标，从而可以对95％的前B ALL患者中的MRD进行定量。这种标准化的实时PCR技术应简化临床试验中对MRD的监测。

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