ESTABLISHMENT OF QUANTITATIVE MEASUREMENT OF FRAGMENTED RED CELLS USING RETICULOCYTE ANALYZER

目的：建立XE-2100全自动血细胞分析仪碎裂红细胞(FRC)定量方法，并探讨其临床应用价值。研究方法：采用加热法制备FRC，建立与手工计数结果呈正相关的新程序。由于缺铁性贫血等红细胞较小的患者出现假阳性数据，还开发了补充门对结果进行修正。目前，XE-2100正在与其他三家医院合作进行临床研究，以评估其对骨髓移植相关血栓性微血管病(BMT-TMA)的诊断效果。结果：1)XE-2100配备的网织红细胞分析仪(RET通道)经荧光染料染色后，通过流式细胞仪分析细胞。在RET通道中，将小红细胞视为FRC，并设置1号门以正向折射手动计数数据(r=0.986)。90例临床标本也呈显著正相关(r=0.876，p&lt；0.001)。然而，从小MCV患者中获得的17例标本由于在1号门检测到正常的小细胞而导致FRC数据假阳性。然后，我们发展了补充的2号门，并得到如下公式。修正的FRC=计算的FRC/e 0.43^&lt；gate2/Total&gt；2)对14例溶血性尿毒症综合征或血栓性血小板减少性紫癜患者进行了该方法的有效性评估。入院时FRC与LD总分无相关性。随着LD同工酶5的升高，我们检验了它们之间的关系(r=0.732，p=0.008)，表明我们的方法检测的FRC反映了溶血。3)骨髓移植患者在感染、VOD等情况下FRC%的增加被检测到。目前，我们正在评估该方法检测BMT-TMA的效果。

Purpose : The purposes were to establish a method of fragmented red cells (FRC) quantification using automated hematology analyzer, XE-2100 and to study about the clinical usefulness of this method.Research ptocess : Experimentary produced FRC by heating were used and new program which shows positive correlation with the results of manual counting has been established. As the patients having small red cells such as iron deficiency anemia shows false pocitive data, complementary gate to revise the results was also developed. Now, clinical study to evaluate its efficacy of the diagnosis of bone marrow transplantation-related thrombotic microangiopathy (BMT-TMA) in collaboration with three other hospitals in Hyogo.Results :1) Reticulocyte analyzer (RET channel) equipped to XE-2100 analyzes cells by flowcytometry after staining with fluorescent dye. In the RET channel, small red cells were considered as FRC, and gate 1 was set to positively refiect the manual counting data (r=0.986). Ninety clinical samples also showed significantly positive correlation between them (r=0.876, p<0.001). However, 17 samples obtained from patients with small MCV had falsely positive FRC data due to detection nomal small cells in gate 1. Then, we have developed complementary gate 2 and the following formula was obtained. Revised FRC=calculated FRC/e 0.43^<gate2/total>2) The usefulness of this method was evaluated in the 14 patients with hemolytic uremic syndrome or thrombotic thrombocytopenic purpura. FRC did not correlated with total LD on admission. As increase in LD isoenzyme 5 was observed, we have checked the relationship between them (r=0.732, p=0.008), indicating that FRC detected by our method reflects hemolysis.3) The increase in FRC % in the patients treated with bone marrow transplantation was detected in several conditions such as infection, VOD and so, on. Now, weare evaluating the efficacy of this method to detect BMT-TMA.