凝血酶-抗凝血酶复合物化学发光免疫分析检测方法的建立
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1.中国科学院深圳先进技术研究院 生物医药与技术研究所 抗体药物研究中心 深圳;2.深圳麦科田生物医疗股份有限公司

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广东省基础与应用基础研究基金委员会 区域联合基金-青年基金项目,2020A1515110840;深圳市科技创新委员会 基础研究(面上项目),JCYJ20190807170801656

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Establishment of chemiluminescence immunoassay method for determination of thrombin-antithrombin complex
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1.Center for Antibody Drug, Institute of Biomedicine and Biotechnology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences;2.Medcaptain Medical Technology Co. Ltd.

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    摘要:

    凝血酶-抗凝血酶复合物(Thrombin antithrombin,TAT)是人体凝血和抗凝血平衡的产物,反映人体的凝血状态,其检测结果可用于血栓性疾病的辅助诊断。该研究建立TAT化学发光免疫分析测定方法,并对其性能进行评估。采用TAT为免疫原制备杂交瘤抗体,以双抗体夹心模式建立TAT化学发光免疫定量检测方法。经过反应的优化,磁珠和吖啶酯抗体的工作浓度确定为 0.20 mg/mL 和 200 ng/mL,样本量为50 μL,磁珠抗体和待测物孵育5 min,再与吖啶酯标记抗体孵育5 min。结果表明,该方法与凝血酶原和抗凝血酶Ⅲ样本没有交叉反应,且与日本希森美康的TAT试剂相关性较高(R>0.99),各项性能指标良好,可满足临床上对血栓性疾病辅助诊断的需求。

    Abstract:

    Thrombin antithrombin (TAT) complex is the product of coagulation and anticoagulation balance. Its detection reflects thrombin status and can be used as an auxiliary in the diagnosis of thrombotic diseases. A chemiluminescence immunoassay method was established to analyze TAT complex in human plasma samples, and its analytical performance were evaluated. The TAT complex was used as immunogen for antibody preparation. TAT measurement method was established using double antibody sandwich format. After the optimization of reaction, the bulk reagent concentrations of magnetic microparticle coated with antibody and acridinium labeled antibody were 0.20 mg/mL and 200 ng/mL, respectively, and the sample size was 50 μL. The reaction of magnetic particle coated antibody and sample were incubated for 5 min, and 5 min for subsequent reacting with acridine ester labeled antibody. There was no cross-reaction with samples containing 0.20 mg/mL prothrombin or 0.31 mg/mL antithrombin Ⅲ. Besides, there was a high correlation (R > 0.99) between this method and TAT test kit of Sysmex. A quantitative chemiluminescence immunoassay method for TAT measurement has been established, and the performance meets customer needs for clinical utility.

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安倩,余枝广,柴凡,等.凝血酶-抗凝血酶复合物化学发光免疫分析检测方法的建立 [J].集成技术,

Citing format
An Qian, Yu Zhiguang, Chai Fan, et al. Establishment of chemiluminescence immunoassay method for determination of thrombin-antithrombin complex[J]. Journal of Integration Technology.

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  • 收稿日期:2023-03-13
  • 最后修改日期:2023-03-13
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  • 在线发布日期: 2023-07-03
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