Quantification of residual disease in chronic myelogenous leukemia patients on interferon-alpha therapy by competitive polymerase chain reaction
Quantification of residual disease in chronic myelogenous leukemia patients on interferon-alpha therapy by competitive polymerase chain reaction
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DOI：
10.1182/blood.v87.4.1549.bloodjournal8741549
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发表时间：
1996-02-15
期刊：
影响因子：
20.3
通讯作者：
Cross, NCP
中科院分区：
文献类型：
--
作者：
Hochhaus, A;Lin, F;Cross, NCP
干扰素-α (IFN-α) 在慢性粒细胞白血病 (CML) 患者中诱导不同程度的细胞遗传学反应。我们试图建立通过竞争性两步逆转录聚合酶链反应 (RT-PCR) 测量的 BCR-ARL 转录本数量与接受 IFN-α 治疗的 CML 患者的细胞遗传学状态之间的关系。研究人员对127名费城染色体阳性(Ph(+))和6名Ph(-)/BCR-ABL(+) CML患者总共250份外周血和55份骨髓样本进行了研究。 21 名患者在诊断时接受了研究，112 名患者在治疗时接受了研究。在接受 IFN-α 治疗的 106 名 Ph(+) 患者中，24 名有完全细胞遗传学缓解，21 名有部分缓解，12 名有轻微缓解，26 名无缓解，23 名未知。使用巢式 RT-PCR，所有 305 个样本的 BCR-ABL 转录本均呈阳性。为了标准化 RNA 和 cDNA 数量和质量变异性的结果，我们量化了每个样品中的总 ABL 转录本作为内部对照。通过与 145 个样品中的葡萄糖-6-磷酸脱氢酶转录物水平进行比较，显示了 ABL 作为内部对照的有效性。完全应答者的中位 BCR-ABL 转录本数量（以及以百分比表示的 BCR-ABL/ABL 比率）为 400/μg RNA（0.04%），部分应答者为 20,500/μg RNA（7.1%），次要应答者为 170,000/μg RNA（21.0%），而无应答者为 430,000/μg RNA（58.7%）。无反应者（P
Interferon-alpha (IFN-alpha) induces cytogenetic responses of variable degree in patients with chronic myelogenous leukemia (CML). We sought to establish the relationship between BCR-ARL transcript numbers measured by competitive 2-step reverse transcription polymerase chain reaction (RT-PCR) and cytogenetic status in CML patients treated with IFN-alpha. A total of 250 peripheral blood and 55 bone marrow samples from 127 Philadelphia chromosome positive (Ph(+)) and 6 Ph(-)/BCR-ABL(+) CML patients were investigated. Twenty-one patients were studied at diagnosis and 112 on treatment. Of the 106 Ph(+) patients treated with IFN-alpha, 24 had a complete cytogenetic response, 21 a partial response, 12 a minor response, 26 no response, and 23 were unknown. Using nested RT-PCR, all 305 samples were positive for BCR-ABL transcripts. To standardize results for variability in RNA and cDNA quantity and quality, we quantified total ABL transcripts in each sample as internal control. The validity of ABL as internal control was shown by comparison with glucose-6-phosphate dehydrogenase transcript levels in 145 samples. The median BCR-ABL transcript numbers (and BCR-ABL/ABL ratios expressed as percentages) were 400/mu g RNA (0.04%) in complete responders, 20,500/mu g RNA (7.1%) in partial responders, 170,000/mu g RNA (21.0%) in minor responders, and 430,000/mu g RNA (58.7%) in nonresponders (P