III. Medizinische Klinik, Universitätsmedizin Mannheim Theodor-Kutzer-Ufer 1-3, 68167 Mannheim, Germany.
Haematologica. 2010 May;95(5):738-44. doi: 10.3324/haematol.2009.016345. Epub 2010 Jan 27.
Rapid identification of diverse fusion genes with involvement of PDGFRA or PDGFRB in eosinophilia-associated myeloproliferative neoplasms is essential for adequate clinical management but is complicated by the multitude and heterogeneity of partner genes and breakpoints.
We established a generic quantitative reverse transcriptase polymerase chain reaction to detect overexpression of the 3'-regions of PDGFRA or PDGFRB as a possible indicator of an underlying fusion.
At diagnosis, all patients with known fusion genes involving PDGFRA (n=5; 51 patients) or PDGFRB (n=5; 7 patients) showed significantly increased normalized expression levels compared to 191 patients with fusion gene-negative eosinophilia or healthy individuals (PDGFRA/ABL: 0.73 versus 0.0066 versus 0.0064, P<0.0001; PDGFRB/ABL: 196 versus 3.8 versus 5.85, P<0.0001). The sensitivity and specificity of the activation screening test were, respectively, 100% and 88.4% for PDGFRA and 100% and 94% for PDGFRB. Furthermore, significant overexpression of PDGFRB was found in a patient with an eosinophilia-associated myeloproliferative neoplasm with uninformative cytogenetics and an excellent response to imatinib. Subsequently, a new SART3-PDGFRB fusion gene was identified by 5'-rapid amplification of cDNA ends polymerase chain reaction (5'-RACE-PCR).
Quantitative reverse transcriptase polymerase chain reaction analysis is a simple and useful adjunct to standard diagnostic assays to detect clinically significant overexpression of PDGFRA and PDGFRB in eosinophilia-associated myeloproliferative neoplasms or related disorders.
在伴嗜酸粒细胞增多的骨髓增生性肿瘤中，快速鉴定涉及 PDGFRA 或 PDGFRB 的多种融合基因对于适当的临床管理至关重要，但由于伙伴基因和断点的多样性和异质性，这变得很复杂。
我们建立了一种通用的定量逆转录聚合酶链反应（qRT-PCR），以检测 PDGFRA 或 PDGFRB 的 3' 区域的过表达，作为潜在融合的可能指标。
在诊断时，所有已知涉及 PDGFRA（n=5；51 例患者）或 PDGFRB（n=5；7 例患者）的融合基因的患者与融合基因阴性嗜酸粒细胞增多症或健康个体相比，均显示出明显增加的归一化表达水平（PDGFRA/ABL：0.73 对 0.0066 对 0.0064，P<0.0001；PDGFRB/ABL：196 对 3.8 对 5.85，P<0.0001）。激活筛选测试的敏感性和特异性分别为 PDGFRA 的 100%和 88.4%，PDGFRB 的 100%和 94%。此外，在一例伴嗜酸粒细胞增多的骨髓增生性肿瘤患者中发现了 PDGFRB 的显著过表达，该患者具有无意义的细胞遗传学和对伊马替尼的良好反应。随后，通过 5'-快速扩增 cDNA 末端聚合酶链反应（5'-RACE-PCR）鉴定了一种新的 SART3-PDGFRB 融合基因。
qRT-PCR 分析是一种简单而有用的辅助标准诊断检测方法，可用于检测伴嗜酸粒细胞增多的骨髓增生性肿瘤或相关疾病中 PDGFRA 和 PDGFRB 的临床显著过表达。
