基于磁性粒子的化学发光免疫分析法的研制及其在血清人降钙素原测定中的应用。
Development of magnetic particle-based chemiluminescence immunoassay for measurement of human procalcitonin in serum.
机构信息
Department of Medical Laboratory, School of Medicine, Hunan Normal University, Changsha 410013, China; Immunodiagnostic Reagents Engineering Research Center of Hunan Province, Hunan Normal University, Changsha 410013, China.
Department of Medical Laboratory, School of Medicine, Hunan Normal University, Changsha 410013, China.
出版信息
J Immunol Methods. 2021 Jan;488:112913. doi: 10.1016/j.jim.2020.112913. Epub 2020 Nov 12.
BACKGROUND
Serum procalcitonin (PCT) has been recognized as a primary biomarker in bacterial infections, and monitoring its concentration could help to evaluate the prognosis of sepsis and guide the antibiotic administration. We aimed to establish a fast and accurate immunoassay for PCT quantitation.
METHODS
Our newly developed monoclonal antibodies (mAbs) against human PCT were preliminarily evaluated by enzyme-linked immunosorbent assay and then used to develop a chemiluminescence enzyme immunoassay (CLEIA). The proposed CLEIA was assessed in analytical performance and applied to measurement of serum PCT.
RESULTS
mAb 2D3 and mAb 8F6 were selected as capture and detection antibody respectively, due to the highest sensitivity for PCT detection with no cross reaction to calcitonin gene-related peptides. The proposed CLEIA based on mAb pair of 2D3/8F6-AP was characterized for a working range from 0.03 to 100 μg/L. An excellent correlation was observed between our proposed assay and the VIDAS BRAHMS PCT assay (r: 0.9825).
CONCLUSION
Our newly developed mAbs and CLEIA can serve as important diagnostic tools for measurement of human PCT in serum.
背景
血清降钙素原（PCT）已被认为是细菌感染的主要生物标志物，监测其浓度有助于评估脓毒症的预后并指导抗生素治疗。我们旨在建立一种快速准确的 PCT 定量免疫分析法。
方法
我们新开发的针对人 PCT 的单克隆抗体（mAb）首先通过酶联免疫吸附试验进行了初步评估，然后用于开发化学发光酶免疫分析（CLEIA）。对所提出的 CLEIA 进行了分析性能评估，并应用于血清 PCT 的测定。
结果
mAb 2D3 和 mAb 8F6 分别被选为捕获抗体和检测抗体，因为它们对 PCT 的检测具有最高的灵敏度，且与降钙素基因相关肽无交叉反应。基于 mAb 对 2D3/8F6-AP 的所提出的 CLEIA 的工作范围为 0.03 至 100μg/L。我们的新方法与 VIDAS BRAHMS PCT 测定法（r：0.9825）之间存在良好的相关性。
结论
我们新开发的 mAb 和 CLEIA 可作为测量血清中人 PCT 的重要诊断工具。
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