Quantitative detection of zeta-chain-associated protein 70 expression in chronic lymphocytic leukemia.
Quantitative detection of zeta-chain-associated protein 70 expression in chronic lymphocytic leukemia.
复制标题
慢性淋巴细胞性白血病中与Zeta-链相关蛋白70表达的定量检测。
DOI：
10.3109/10428194.2012.715349
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发表时间：
2013-03
影响因子：
2.6
通讯作者：
Tang CM
中科院分区：
文献类型：
--
作者：
Zhu P;Degheidy HA;Marti GE;Li S;Abbasi F;Wiestner A;Amstutz P;Tang CM
zeta链相关蛋白70（ZAP-70）的过表达最近被认为是慢性淋巴细胞白血病（CLL）侵袭性形式的独立预后标志物。本研究的目的是证明定量检测B细胞中ZAP-70蛋白的可行性和实施，以明确区分CLL患者与疾病的侵袭性形式。从患者血液中分离B细胞并裂解。使用免疫磁性荧光测定法检测释放的ZAP-70蛋白。使用Jurkat细胞和重组ZAP-70（rZAP-70）开发测定方案。检测限被确定为低于125个Jurkat细胞和39 pg的rZAP-70蛋白。信号响应在宽的动态范围内是线性的，从125到40 000 Jurkat细胞/测试（R2 = 0.9987）和从0到40 000 pg rZAP-70蛋白/测试（R2 = 0.9928）。20例CLL患者的结果与流式细胞术分析密切相关。两种方法阳性和阴性结果的一致性分别为100%（7/7）和92%（12/13），而两种方法之间的总体一致性为95%。本文报道的测定方法是一种简单、可靠和可重复的方法，用于定量检测患者白血病细胞中的ZAP-70，而不需要细胞固定或透化。ZAP-70信号在宽的动态范围内呈线性，我们认为这使得能够定量评估疾病过程中ZAP-70表达的微小变化以及对治疗干预的反应。
Overexpression of zeta-chain-associated protein 70 (ZAP-70) was recently recognized as an independent prognostic marker for the aggressive form of chronic lymphocytic leukemia (CLL). The objective of this study was to demonstrate the feasibility and implementation of quantitative detection of ZAP-70 protein in B cells to clearly distinguish patients with CLL with the aggressive form of the disease. B cells were isolated from patient blood and lysed. Released ZAP-70 protein was detected using an immunomagnetic fluorescence assay. The assay protocol was developed using Jurkat cells and recombinant ZAP-70 (rZAP-70). The limit of detection was determined to be lower than 125 Jurkat cells and 39 pg of rZAP-70 protein. The signal response was linear over a wide dynamic range, from 125 to 40 000 Jurkat cells per test (R2 = 0.9987) and from 0 to 40 000 pg rZAP-70 protein per test (R2 = 0.9928). The results from 20 patients with CLL correlated strongly with flow cytometry analysis. Concordance between the two methods for positive and negative results was 100% (7/7) and 92% (12/13), respectively, while the overall concordance between the two methods was 95%. The assay reported here is a simple, reliable and reproducible method for quantitative detection of ZAP-70 in patient leukemic cells, without the need for cell fixation or permeabilization. The ZAP-70 signal was linear over a wide dynamic range, which we believe enables quantitative assessment of small changes in ZAP-70 expression over the course of the disease and in response to therapeutic intervention.
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影响因子：
20.1
作者：
Pryshchep S;Goronzy JJ;Parashar S;Weyand CM
通讯作者：
Weyand CM
DOI：
10.1073/pnas.0404432101
发表时间：
2004-08-10
影响因子：
11.1
作者：
Calin, GA;Liu, CG;Croce, CM
通讯作者：
Croce, CM
影响因子：
158.5
作者：
Rassenti, LZ;Huynh, L;Kipps, TJ
通讯作者：
Kipps, TJ
影响因子：
158.5
作者：
Crespo, M;Bosch, F;Montserrat, E
通讯作者：
Montserrat, E
影响因子：
9.3
作者：
Stamatopoulos, Basile;Meuleman, Nathalie;Lagneaux, Laurence
通讯作者：
Lagneaux, Laurence