Poopak Behzad, Valeshabad Ali Kord, Elahi Fazel, Rezvani Hamid, Khosravipour Gelareh, Jahangirpour Mohammad Ali, Bolouri Shirin, Golkar Tolou, Salari Fatemeh, Shahjahani Mohammad, Saki Najmaldin
Tehran Medical Branch, Islamic Azad University, Zargandeh St, Shariati Ave, Tehran, Iran.
Students' Scientific Research Center (SSRC), Tehran University of Medical Sciences (TUMS), Tehran, Iran ; National Foundation of Elites, Tehran, Iran.
Indian J Hematol Blood Transfus. 2015 Mar;31(1):38-45. doi: 10.1007/s12288-014-0387-z. Epub 2014 May 4.
This study investigates PCR analysis of immunoglobulin heavy chain (IgH) and T cell receptor (TCR) gene rearrangements on paraffin-embedded tissue sections and bone marrow aspirates of patients suspected to have lymphoproliferative disorders but with inconclusive diagnosis in histopathological examination. 130 samples of patients with inconclusive immunohistochemistry results were evaluated for clonal rearrangement of IgH and TCR genes. Based on histopathology examination, the patients were divided into three groups: the first group without any definite diagnosis of lymphoproliferative disorders (60 cases, 46.2 %), the second group suspected to have a lymphoproliferative disorder but in favor of benign disorders (19 cases, 14.6 %) and the third group suspect to lymphoproliferative disorders but relatively in favor of malignant disorders (51 cases, 39.2 %). After DNA extraction and quality control, semi-nested PCR was performed using consensus primers for amplification of TCR-γ and CDR-3 regions of IgH genes. PCR products were analyzed after heteroduplex analysis using polyacrylamide gel electrophoresis, and were subject to silver staining. Totally, in over half of the cases (55.4 %), a monoclonal pattern was found in IgH or TCR-γ genes rearrangements. Monoclonal IgH gene rearrangement was detected in 48.1 % of patients, whereas monoclonal TCR-γ gene rearrangement was found in 33.6 % of them, which was not statistically significant (P = 0.008). Only in 32 patients (24.6 %) were the results of TCR-γ and IgH gene rearrangements consistent with respect to the presence (2.3 %) or absence (22.3 %) of monoclonality. Finally, PCR analysis of TCR-γ and IgH gene rearrangements led to definite diagnosis in 105 patients (80.8 %), and only 25 cases (19.2 %) remained inconclusive. Our results emphasize the usefulness of gene rearrangement study in cases without a definite diagnosis in immunohistochemistry studies. Multiple PCR analysis results when combined with patient's clinical course and immunohistochemistry can lead to early diagnosis and subsequent therapy.
本研究调查了对疑似患有淋巴增殖性疾病但组织病理学检查诊断不明确的患者石蜡包埋组织切片和骨髓穿刺液进行免疫球蛋白重链（IgH）和T细胞受体（TCR）基因重排的PCR分析。对130例免疫组化结果不明确的患者样本进行了IgH和TCR基因的克隆重排评估。根据组织病理学检查，将患者分为三组：第一组未明确诊断为淋巴增殖性疾病（60例，46.2%），第二组疑似患有淋巴增殖性疾病但倾向于良性疾病（19例，14.6%），第三组疑似患有淋巴增殖性疾病但相对倾向于恶性疾病（51例，39.2%）。DNA提取和质量控制后，使用共有引物进行半巢式PCR，以扩增IgH基因的TCR-γ和CDR-3区域。PCR产物经聚丙烯酰胺凝胶电泳进行异源双链分析后进行分析，并进行银染。总共，超过一半的病例（55.4%）在IgH或TCR-γ基因重排中发现单克隆模式。48.1%的患者检测到单克隆IgH基因重排，而33.6%的患者发现单克隆TCR-γ基因重排，差异无统计学意义（P = 0.008）。仅32例患者（24.6%）的TCR-γ和IgH基因重排结果在单克隆性存在（2.3%）或不存在（22.3%）方面一致。最后，TCR-γ和IgH基因重排的PCR分析导致105例患者（80.8%）明确诊断，仅25例（19.2%）仍诊断不明确。我们的结果强调了基因重排研究在免疫组化研究中未明确诊断病例中的有用性。多种PCR分析结果与患者的临床病程和免疫组化相结合可导致早期诊断及后续治疗。
