Flow cytometric immunobead assay for quantitative detection of platelet autoantibodies in immune thrombocytopenia patients.
Flow cytometric immunobead assay for quantitative detection of platelet autoantibodies in immune thrombocytopenia patients.
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流式细胞术免疫珠法定量检测免疫性血小板减少症患者血小板自身抗体
DOI：
10.1186/s12967-017-1317-2
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发表时间：
2017-10-23
影响因子：
7.4
通讯作者：
He Y
中科院分区：
文献类型：
--
作者：
Zhai J;Ding M;Yang T;Zuo B;Weng Z;Zhao Y;He J;Wu Q;Ruan C;He Y
背景：血小板自身抗体检测对免疫性血小板减少症（ITP）的诊断和预后至关重要。因此，我们旨在建立一种用于ITP血小板自身抗体评估的定量流式细胞免疫珠法（FCIA）。方法采用捕集微珠偶联抗gpix、-GPIb、-GPIIb、-GPIIIa和p -选择素抗体，结合250例ITP患者、163例非ITP患者和243例健康对照血浆样本产生的血小板结合自身抗体复合物，以异硫氰酸荧光素（FITC）偶联二抗为检测试剂，流式细胞术记录平均荧光强度（MFI）信号。定量FCIA测定法的测定内和测定间变化进行了评估。比较定量与定性FCIA法和单克隆抗体固定化血小板抗原（MAIPA）法的特异性、敏感性和准确性。最后，通过定量FCIA监测8例新诊断ITPs的治疗过程。结果FCIA定量检测抗gpix、-GPIb、-GPIIIa、-GPIIb和- p -选择素自身抗体的变异系数（CV）分别为9.4、3.8、5.4、5.1和5.8%。与非ITP患者或健康对照相比，我们的定量FCIA检测到ITP患者抗血小板糖蛋白GPIX、GPIb、GPIIIa、GPIIb和p -选择素的自身抗体水平升高。联合5种自身抗体时，定量检测的灵敏度、特异性和准确性分别为73.13、81.98和78.65%，MAIPA检测的灵敏度、特异性和准确性分别为41.46、90.41和72.81%。结论建立了FCIA定量检测方法。血小板自身抗体水平的降低可以通过我们的定量FCIA在ITP患者接受皮质类固醇治疗后得到证实。本方法不仅对ITP的诊断有益，而且对ITP的治疗监测也有重要意义。
BackgroundPlatelet autoantibody detection is critical for immune thrombocytopenia (ITP) diagnosis and prognosis. Therefore, we aimed to establish a quantitative flow cytometric immunobead assay (FCIA) for ITP platelet autoantibodies evaluation.MethodsCapture microbeads coupled with anti-GPIX, -GPIb, -GPIIb, -GPIIIa and P-selectin antibodies were used to bind the platelet-bound autoantibodies complex generated from plasma samples of 250 ITP patients, 163 non-ITP patients and 243 healthy controls, a fluorescein isothiocyanate (FITC)-conjugated secondary antibody was the detector reagent and mean fluorescence intensity (MFI) signals were recorded by flow cytometry. Intra- and inter-assay variations of the quantitative FCIA assay were assessed. Comparisons of the specificity, sensitivity and accuracy between quantitative and qualitative FCIA or monoclonal antibody immobilization of platelet antigen (MAIPA) assay were performed. Finally, treatment process was monitored by our quantitative FCIA in 8 newly diagnosed ITPs.ResultsThe coefficient of variations (CV) of the quantitative FCIA assay were respectively 9.4, 3.8, 5.4, 5.1 and 5.8% for anti-GPIX, -GPIb, -GPIIIa, -GPIIb and -P-selectin autoantibodies. Elevated levels of autoantibodies against platelet glycoproteins GPIX, GPIb, GPIIIa, GPIIb and P-selectin were detected by our quantitative FCIA in ITP patients compared to non-ITP patients or healthy controls. The sensitivity, specificity and accuracy of our quantitative assay were respectively 73.13, 81.98 and 78.65% when combining all 5 autoantibodies, while the sensitivity, specificity and accuracy of MAIPA assay were respectively 41.46, 90.41 and 72.81%.ConclusionsA quantitative FCIA assay was established. Reduced levels of platelet autoantibodies could be confirmed by our quantitative FCIA in ITP patients after corticosteroid treatment. Our quantitative assay is not only good for ITP diagnosis but also for ITP treatment monitoring.
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影响因子：
6.5
作者：
Arnold, Donald M.;Vrbensky, John R.;Nazy, Ishac
通讯作者：
Nazy, Ishac
影响因子：
20.3
作者：
Rodeghiero, Francesco;Stasi, Roberto;George, James N.
通讯作者：
George, James N.
影响因子：
10.4
作者：
McMillan, R;Wang, L;Tani, P
通讯作者：
Tani, P
DOI：
10.1073/pnas.0631579100
发表时间：
2003-05-13
影响因子：
11.1
作者：
Schuck, S;Honsho, M;Simons, K
通讯作者：
Simons, K
影响因子：
12.8
作者：
FREEDMAN, J;GARVEY, MB;BLANCHETTE, V
通讯作者：
BLANCHETTE, V