Paro Mirjana Mariana Kardum, Siftar Zoran, Kardum-Skelin Ika, Sustercić Dunja, Nazor Aida, Flegar-Mestrić Zlata, Jaksić Branimir
Institute of Clinical Chemistry, "Merkur" University Hospital, Zagreb, Croatia.
Coll Antropol. 2010 Jun;34(2):359-65.
Flow cytometry immunophenotyping (FCI) has an important role in the clinic work-up of fine needle aspirates (FNAs) of lymph nodes. Its standardization has been defined by proposed analytical protocols and procedures used to assure proper analytical results also in those non-routine samples. In Institute of Clinical Chemistry, "Merkur" University Hospital, FCI is accredited method according to laboratory accreditation standard ISO 15189. According to this laboratory accreditation standard, participation in external quality assessment (EQA) programs is a prerequisite for assuring integrity and quality of the entire laboratory process. A critical analysis of our institutional experience in the feasibility of FCI of the material obtained by FNA of lymph nodes with suspected lymphoma represented the purpose of the study. During an eight-year period in Institute of Clinical Chemistry, "Merkur" University Hospital, a total of 1295 FNA analysis was done, 245 of them with a possible diagnosis of B-cell Non-Hodgkin lymphomas (B-NHL) formed the basis of the study. Lymphocytes were isolated on density gradient according to Boyum et al. The average feasibility of FNAs for FCI analysis was 86% (ranged 78-93%). An acceptable total cell number in FNAs for FCI analysis (4257) was established. In total population of respondents statistical significances in expressions of cellular antigens CD3, CD5, CD22, CD23, CD19 and CD5 on B-cells (CD5+CD19+) between patient's with final diagnosis of benign, reactive lymphoid proliferations and patient's with diagnosis of B-NHL were found. EQA results analysis showed that all results were either inside target values (X +/- 1SD) or inside accepted values (X +/- 2SD). Compatibility of the restriction of immunoglobulins light chains determinated by FCI and cytomorphology diagnosis depends on the choice of criterion values of the light chains ratio which determine the monoclonality. According to the matrix of shares of all classified data of retained neural network, ranges of diagnostic sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and prevalency of 82%, 72%, 93%, 48%, and 72% were produced. As a conclusion, FCI is a reliable methodology for phenotyping FNAs of lymph nodes with suspected B-NHLs detecting their clonality easily.
流式细胞术免疫表型分析（FCI）在淋巴结细针穿刺抽吸物（FNA）的临床检查中具有重要作用。其标准化已通过提议的分析方案和程序来定义，这些方案和程序用于确保在那些非常规样本中也能获得恰当的分析结果。在“梅尔库”大学医院临床化学研究所，根据实验室认可标准ISO 15189，FCI是一项认可的方法。根据该实验室认可标准，参与外部质量评估（EQA）计划是确保整个实验室过程完整性和质量的前提条件。对我们机构在对疑似淋巴瘤的淋巴结FNA所获材料进行FCI可行性方面的经验进行批判性分析是本研究的目的。在“梅尔库”大学医院临床化学研究所的八年期间，共进行了1295次FNA分析，其中245次可能诊断为B细胞非霍奇金淋巴瘤（B-NHL），构成了本研究的基础。根据博伊姆等人的方法，通过密度梯度分离淋巴细胞。FNA用于FCI分析的平均可行性为86%（范围为78%-93%）。确定了FCI分析中FNA可接受的总细胞数（4257）。在全部受试人群中，最终诊断为良性、反应性淋巴组织增生的患者与诊断为B-NHL的患者在B细胞（CD5+CD19+）上细胞抗原CD3、CD5、CD22、CD23、CD19和CD5的表达存在统计学意义。EQA结果分析表明，所有结果要么在靶值（X +/- 1SD）范围内，要么在可接受值（X +/- 2SD）范围内。FCI测定的免疫球蛋白轻链限制与细胞形态学诊断的兼容性取决于确定单克隆性的轻链比例标准值的选择。根据保留神经网络的所有分类数据的份额矩阵，得出诊断敏感性、特异性、阳性预测值（PPV）、阴性预测值（NPV）和患病率的范围分别为82%、72%、93%、48%和72%。结论是，FCI是一种可靠的方法，可用于对疑似B-NHL的淋巴结FNA进行表型分析，并能轻松检测其克隆性。
