Detecting Minimal Residual Disease in Neuroblastoma: The Superiority of a Panel of Real-Time Quantitative PCR Markers
Detecting Minimal Residual Disease in Neuroblastoma: The Superiority of a Panel of Real-Time Quantitative PCR Markers
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DOI：
10.1373/clinchem.2008.117945
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发表时间：
2009-07-01
影响因子：
9.3
通讯作者：
Tytgat, Godelieve A. M.
中科院分区：
文献类型：
--
作者：
Stutterheim, Janine;Gerritsen, Annemieke;Tytgat, Godelieve A. M.
背景：基于 PCR 的神经母细胞瘤 (NB) 患者微小残留病 (MRD) 检测可用于初始分期和监测骨髓 (BM) 和外周血 (PB) 的治疗反应。 PHOX2B已被确定为敏感且特异的MRD标记物；然而，其表达因肿瘤而异。因此，一组标记物可以提高敏感性。 方法：为了识别 NB 的其他 MRD 标记物，我们通过比较健康组织和 NB 组织的 SAGE（基因表达系列分析）文库，然后对肿瘤 (n = 56)、对照 BM (n = 51)、PB (n = 37) 和细胞亚群样本进行广泛的实时定量 PCR (RQ-PCR) 测试来选择基因。在来自 82 名荷兰 4 期 NB 患者的 222 个 NB 样本中确定了一组附加值（54 个诊断 BM 样本、143 个治疗期间/治疗后的 BM 样本和 25 个 PB 样本）。 结果：我们确定了 2 个用于 NB 患者 MRD 检测的特异性 RQ-PCR 标记物组：I 用于分析 BM 样本（PHOX2B、TH、DDC、CHRNA3 和 GAP43），I 用于分析 PB 样本（PHOX2B、TH、DDC、DBH 和 CHRNA3）。这些标记物在 NB 肿瘤中均表现出高表达，而在对照 BM 或 PB 样本中则无或低表达。在患者样本中，PHOX2B 标记检测到大多数阳性样本。然而，在 PB 样本中，7 个 PHOX2B 阴性样本中的 3 个对 1 个或多个标记物呈阳性，并且在治疗期间的 BM 检查中，7%（86 个中的 6 个）PHOX2B 阴性样本对另一种标记物呈阳性。 结论：由于 BM 和 PB 中标记物的敏感性存在差异，我们建议使用 2 个不同的面板来检测这些区室中的 MRD。 (C) 2009 美国临床化学协会
BACKGROUND: PCR-based detection of minimal residual disease (MRD) in neuroblastoma (NB) patients can be used for initial staging and monitoring therapy response in bone marrow (BM) and peripheral blood (PB). PHOX2B has been identified as a sensitive and specific MRD marker; however, its expression varies between tumors. Therefore, a panel of markers could increase sensitivity.METHODS: To identify additional MRD markers for NB, we selected genes by comparing SAGE (serial analysis of gene expression) libraries of healthy and NB tissues followed by extensive real-time quantitative PCR (RQ-PCR) testing in samples of tumors (n = 56), control BM (n = 51), PB (n = 37), and cell subsets. The additional value of a panel was determined in 222 NB samples from 82 Dutch stage 4 NB patients (54 diagnosis BM samples, 143 BM samples during/after treatment, and 25 PB samples).RESULTS: We identified 2 panels of specific RQ-PCR markers for MRD detection in NB patients: I for analysis of BM samples (PHOX2B, TH, DDC, CHRNA3, and GAP43) and I for analysis of PB samples (PHOX2B, TH, DDC, DBH, and CHRNA3). These markers all showed high expression in NB tumors and no or low expression in control BM or PB samples. In patients' samples, the PHOX2B marker detected most positive samples. In PB samples, however, 3 of 7 PHOX2B-negative samples were positive for 1 or more markers, and in BM examinations during treatment, 7% (6 of 86) of the PHOX2B-negative samples were positive for another marker.CONCLUSIONS: Because of differences in the sensitivities of the markers in BM and PB, we advise the use of 2 different panels to detect MRD in these compartments. (C) 2009 American Association for Clinical Chemistry