Peluso A L, Cozzolino I, Bottiglieri A, Lucchese L, Di Crescenzo R M, Langella M, Selleri C, Zeppa P
Anatomia Patologica, University of Salerno, Salerno, Italy.
DEA, University "G. Marconi" of Rome, Rome.
Cytopathology. 2017 Jun;28(3):203-215. doi: 10.1111/cyt.12402. Epub 2016 Dec 22.
To evaluate and compare the DNA yield and quality extracted from lymph node fine needle cytology (FNC) samples stored on FTA cards to those cryopreserved, and to assess the immunoglobulin heavy and light chains (IGHK) and T-Cell receptor beta and gamma chains (TCRBG) PCR tests.
DNA extractions were performed on FNC of 80 non-Hodgkin lymphomas (NHL), four myelomas and 56 benign reactive hyperplasias (BRH) cryopreserved and stored on FTA cards. The JAK2 gene was amplified to assess the DNA integrity and the IGHK/TCRBG clonality status was tested.
IGHK monoclonality was found in 99% of B-cell NHL and 100% of myeloma. TCRBG monoclonality was found in 100% of T-cell NHL. TCRBG polyclonality was detected in 97% of B-cell NHL, 100% of myeloma and 96% of BRH. IGHK/TCRBG PCR data were confirmed by histological and/or follow-up controls. No differences were found in the DNA quality between cryopreservation and FTA cards storage methods.
IGHK/TCRBG PCR of the lymphoproliferative process on FTA cards is comparable to those cryopreserved. FTA cards can be used to store lymph node FNC for further molecular investigations.
评估并比较储存在FTA卡上的淋巴结细针穿刺细胞学(FNC)样本与冷冻保存样本的DNA产量和质量,并评估免疫球蛋白重链和轻链(IGHK)以及T细胞受体β链和γ链(TCRBG)的聚合酶链反应(PCR)检测。
对80例非霍奇金淋巴瘤(NHL)、4例骨髓瘤和56例良性反应性增生(BRH)的FNC样本进行DNA提取,这些样本分别进行了冷冻保存和储存在FTA卡上。扩增JAK2基因以评估DNA完整性,并检测IGHK/TCRBG的克隆性状态。
在99%的B细胞NHL和100%的骨髓瘤中发现IGHK单克隆性。在100%的T细胞NHL中发现TCRBG单克隆性。在97%的B细胞NHL、100%的骨髓瘤和96%的BRH中检测到TCRBG多克隆性。IGHK/TCRBG PCR数据经组织学和/或随访对照证实。冷冻保存和FTA卡储存方法之间的DNA质量没有差异。
FTA卡上淋巴增殖过程的IGHK/TCRBG PCR与冷冻保存样本的结果相当。FTA卡可用于储存淋巴结FNC样本以供进一步分子研究。