Svachova Michaela, Tichy Martin, Flodr Patrik, Steigerova Jana, Kolar Zdenek, Bouchal Jan
Department of Clinical and Molecular Pathology, Faculty of Medicine and Dentistry, Palacky University Olomouc and University Hospital Olomouc, Czech Republic.
Institute of Molecular and Translation Medicine, Faculty of Medicine and Dentistry, Palacky University Olomouc, Czech Republic.
Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2017 Jun;161(2):197-205. doi: 10.5507/bp.2017.006. Epub 2017 Mar 14.
Lymphoproliferative disease often presents the clinician and pathologist with a diagnostic dilemma, particularly in the early course of the disease.
We used modified BIOMED-2 protocols to detect monoclonal expansions of immunoglobulin heavy chain (IgH) and T-cell receptor (TCR) genes in 957 formalin-fixed paraffin-embedded samples from 717 patients. To eliminate false-positive results, heteroduplex analysis was used after PCR reactions. The impact of different fixatives on DNA quality and performance of PCR was assessed.
In the class of B lymphomas we detected clonal IgH rearrangement in nearly 80% of cases and in the class of T lymphomas in 64% of cases. Performance of the assays was 94.7% and 92.5% for IgH and TCR clonality, respectively. Clonality rates in various B and T lymphomas were in concordance with previous studies. We also present 10 difficult cases where PCR analysis of IgH and TCR gene rearrangements significantly contributed to a decision on the correct diagnosis.
These results confirm that the PCR-based analysis is suitable as a routine method and is helpful in establishing a diagnosis in morphologically unclear cases.
淋巴增殖性疾病常常给临床医生和病理学家带来诊断难题，尤其是在疾病早期。
我们采用改良的BIOMED-2方案，对717例患者的957份福尔马林固定石蜡包埋样本中的免疫球蛋白重链（IgH）和T细胞受体（TCR）基因的单克隆扩增进行检测。为消除假阳性结果，在PCR反应后采用异源双链分析。评估了不同固定剂对DNA质量和PCR性能的影响。
在B淋巴瘤类别中，我们在近80%的病例中检测到克隆性IgH重排，在T淋巴瘤类别中为64%。IgH和TCR克隆性检测的成功率分别为94.7%和92.5%。各种B和T淋巴瘤的克隆率与先前研究一致。我们还展示了10例疑难病例，其中IgH和TCR基因重排的PCR分析对做出正确诊断的决策有显著帮助。
这些结果证实基于PCR的分析适合作为常规方法，有助于在形态学不明确的病例中确立诊断。
