Establishment of a novel homogeneous nanoparticle-based assay for sensitive procalcitonin detection of ultra low-volume serum samples
Establishment of a novel homogeneous nanoparticle-based assay for sensitive procalcitonin detection of ultra low-volume serum samples
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建立一种新型均质纳米颗粒检测方法，用于超低体积血清样品的降钙素原的灵敏检测
DOI：
10.2147/ijn.s173776
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发表时间：
2018-01-01
影响因子：
8
通讯作者：
Liu，Tiancai
中科院分区：
文献类型：
--
作者：
Li，Peng;Chen，Zhenhua;Liu，Tiancai
脓毒症是一种潜在的致命性全身感染，在世界范围内具有显著的死亡率。虽然C反应蛋白（CRP）、白细胞介素-6（IL-6）和降钙素原（PCT）可能是脓毒症诊断的生物标志物，但PCT比CRP或IL-6更敏感和特异。我们的目的是建立一种有效的免疫分析，准确地检测PCT在人血清中的早期诊断脓毒症。材料与方法我们建立了一种新的扩增发光邻近均相检测法（AlphaLISA），用于血清中PCT的定量检测。在该测定中，使用一对抗体捕获血清中的PCT，并在37°C下孵育15分钟后形成夹心复合物。结果PCT浓度在0.016 ~ 100 ng/mL范围内线性关系良好。检测限为18.6 pg/mL。结果表明，该方法的重现性、回收率和特异性均符合临床检测要求。与市售酶联荧光法（ELFA）试剂盒的决定系数（R2）为0.93045。结论该方法灵敏度高、动态范围宽，可用于PCT检测。与传统的异质性检测方法如ELISA相比，该检测方法测量了PCT的同质形式的浓度，并在较短的检测时间内提供了更准确的结果。我们期望这种新的方法将有助于脓毒症患者的早期筛查和预后评估。
Purpose Sepsis is a potentially fatal systemic body infection with a significant mortality rate worldwide. Although C-reactive protein (CRP), interleukin-6 (IL-6), and procalcitonin (PCT) might be biomarkers for sepsis diagnosis, PCT is more sensitive and specific than CRP or IL-6. We aimed to establish an efficient immunoassay that precisely detects PCT in human serum for the early diagnosis of sepsis. Materials and methods We developed a novel amplified luminescent proximity homogeneous assay (AlphaLISA) for the quantitative detection of PCT in serum. In this assay, a pair of antibodies was used to capture PCT in serum and to form sandwich complexes after incubating for 15 minutes at 37°C. Results PCT concentrations were determined within a linear range of 0.016–100 ng/mL. The limit of detection was 18.6 pg/mL. The results demonstrate that the reproducibility, recovery, and specificity of this assay for PCT meet the requirements of clinical detection. The coefficient of determination (R2) between this method and commercially available enzyme-linked fluorescent assay (ELFA) kits was estimated to be 0.93045 in clinical serum testing. Conclusion The novel assay for PCT detection was robust with high sensitivity and a broad dynamic range. Compared with conventional heterogeneous detection methods such as ELISA, this assay measured the concentration of the homogeneous form of PCT and provided results that are more accurate within a shorter detection time. We expect that this novel method will be useful for the early screening and prognosis evaluation of patients with sepsis.