Laboratory evaluation of a flow cytometric BCR-ABL immunobead assay
Laboratory evaluation of a flow cytometric BCR-ABL immunobead assay
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流式细胞术 BCR-ABL 免疫珠测定的实验室评估
DOI：
10.1515/cclm.2011.834
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
J. Kappelmayer
中科院分区：
文献类型：
--
作者：
Z. Hevessy;R. Hudák;Valéria Kiss;P. Antal;M. Udvardy;L. Rejtő;L. Szerafin;J. Kappelmayer
摘要背景：一种新的流式细胞术（FC）BCR-ABL免疫珠检测已被开发出来。在这里，我们提出了市售试剂盒的实验室评价。方法：按照生产商的说明书分离、裂解和处理单核细胞。吸附至捕获珠的抗BCR抗体结合裂解细胞的BCR-ABL融合蛋白，藻红蛋白（PE）结合的抗ABL抗体为检测试剂，并通过流式细胞术记录平均荧光强度（MFI）信号。用定量PCR检测t（9;22）（q34;q11）易位。结果：20例正常人外周血MFI值为88±8（均数±标准差），变异系数为9%。K562细胞用作阳性对照。批内不精密度极佳（正常范围为3.7%，病理范围为10%）。截止值选择在MFI 112，灵敏度和特异性均为100%。共分析了17例慢性粒细胞白血病（CML）和16例急性白血病样本。所有PCR阳性样本（n=14）均为FC法阳性，阴性结果也一致（n=15）。冷冻的细胞裂解物可以储存长达4周，而MFI信号没有显著降低。结论：FC BCR-ABL检测是一种快速、可重复和可靠的方法，可纳入标准流式细胞术方案，以帮助临床决策。
Abstract Background: A new flow cytometric (FC) BCR-ABL immunobead assay has been developed recently. Here we present the laboratory evaluation of the commercially available kit. Methods: Mononuclear cells were isolated, lysed and processed according to the instructions of the manufacturer. Anti-BCR antibodies adsorbed to capture beads bind the BCR-ABL fusion proteins of the lysed cells, a phycoerythrin (PE)-conjugated anti-ABL antibody is the detector reagent and mean fluorescence intensity (MFI) signals were recorded by flow cytometry. Detection of t(9;22)(q34;q11) translocation was carried out with a quantitative PCR assay. Results: MFI results of 20 normal peripheral blood samples were 88±8 (mean±SD), CV 9%. K562 cells were used as positive control. Within-batch imprecision was excellent (3.7% in the normal and 10% in the pathological range). Cut-off was chosen at MFI 112, where both sensitivity and specificity were 100%. Altogether 17 chronic myeloid leukemia (CML) and 16 acute leukemia samples were analyzed. All PCR positive samples (n=14) were positive with the FC method and negative results were also concordant (n=15). Frozen cell lysates can be stored up to 4 weeks without significant decrease of MFI signal. Conclusions: The FC BCR-ABL assay is a fast, reproducible and reliable method that may be incorporated into standard flow cytometric protocols to help clinical decision-making.
影响因子：
158.5
作者：
Druker, BJ;Talpaz, M;Sawyers, CL
通讯作者：
Sawyers, CL