Biochemia Medica:分析前经典和替代补体途径活性的损失如何?

2019-11-03 MedSci MedSci原创

补体功能分析能够为整个补体反应级联的完整性提供信息。对于调查可疑的补体缺陷,这些检测尤为重要。个别补体成分分析前的不稳定性可能会导致检测结果的错误性降低。为了为这种或潜在的替代实践提供理论依据,本研究旨在扩展广泛使用补体系统的测试的分析前稳定性方面的知识。

补体功能分析能够为整个补体反应级联的完整性提供信息。对于调查可疑的补体缺陷,这些检测尤为重要。个别补体成分分析前的不稳定性可能会导致检测结果的错误性降低。为了为这种或潜在的替代实践提供理论依据,本研究旨在扩展广泛使用补体系统的测试的分析前稳定性方面的知识。我们评估了时间、温度和EDTA对经典(CH50)和替代途径(AP50)功能检测结果的影响。

我们使用浊度(C3d)免疫固定(C3c)技术来进行基础补体系统活性测试分析前阶段的调查。采用溶血活性测定法和C5b-9新表位elisa法分别对经典和替代途径功能进行定量测定。对5名健康志愿者的血液进行采样,并在不同的条件下允许补体成分存在一定的降解。

在冰上孵育的血清样本中,CH50AP50稳定保持约一周。与室温下的血清相比,EDTA血浆中的CH50活性下降速度几乎是前者的两倍。相反,与血清相比,EDTA血浆中AP50活性降低的速度是室温下的两倍。

功能补体分析的首选标本仍然是冰上的血清。在没有冰运血清的情况下,室温(不超过24小时)保存的血清适用于经典和替代途径分析。对于其他途径分析,EDTA的稳定作用允许EDTA血浆的延长使用(不超过4)。在这些情况下,至少85%的基线补体活性仍然存在。

原始出处:

Koen O.A. VercauterenStijn LambrechtPreanalytical classical and alternative complement pathway activity loss

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    2020-07-21 jeanqiuqiu
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