Abstract：
OBJECTIVE: It has been reported that serum Clara cell secreted protein 16 (CC16) is a potential biomarker for lung injury diseases, but currently, there is no other method that is faster, more accurate, or more sensitive being applied in clinical practice apart from ELISA. The current study was designed to established a magnetic nanoparticles chemiluminescence immunoassay (MNPs-CLIA) for highly sensitive automated detection of serum Clara cell secretory protein 16 (CC16), and validated its diagnostic performance for lung disease.
METHODS: The study included the expression of CC16 recombinant protein, the preparation and screening of its monoclonal antibody (MAb), as well as the construction, optimization and analytical evaluation of the MNPs-CLIA method. The clinical application value of this method was investigated by detecting CC16 level in 296 serum samples.
RESULTS: The linear range of the MNPs-CLIA assay system was 0.2-50 ng/mL, and the limit of detection was 0.037 ng/mL. Performance parameters such as specificity, recovery rate, and precision can meet the industry standards of in vitro diagnostic reagents. The established method reveals consistent results with ELISA (R2=0.9962) currently used clinically, and it also exhibits satisfactory diagnostic efficacy of silicosis, chronic obstructive pulmonary disease (COPD), and pulmonary sarcoidosis, with areas under the curve (AUC) of 0.9748, 0.8428 and 0.9128, respectively.
CONCLUSIONS: Our established MNPs-CLIA method has the advantages of automation, high throughput, rapidity, and simplicity, and can be promoted for widely popularized in clinical applications. MNPs-CLIA detection of serum CC16 has efficient diagnostic potentiality for predicting and diagnosing lung diseases.
METHODS: The study included the expression of CC16 recombinant protein, the preparation and screening of its monoclonal antibody (MAb), as well as the construction, optimization and analytical evaluation of the MNPs-CLIA method. The clinical application value of this method was investigated by detecting CC16 level in 296 serum samples.
RESULTS: The linear range of the MNPs-CLIA assay system was 0.2-50 ng/mL, and the limit of detection was 0.037 ng/mL. Performance parameters such as specificity, recovery rate, and precision can meet the industry standards of in vitro diagnostic reagents. The established method reveals consistent results with ELISA (R2=0.9962) currently used clinically, and it also exhibits satisfactory diagnostic efficacy of silicosis, chronic obstructive pulmonary disease (COPD), and pulmonary sarcoidosis, with areas under the curve (AUC) of 0.9748, 0.8428 and 0.9128, respectively.
CONCLUSIONS: Our established MNPs-CLIA method has the advantages of automation, high throughput, rapidity, and simplicity, and can be promoted for widely popularized in clinical applications. MNPs-CLIA detection of serum CC16 has efficient diagnostic potentiality for predicting and diagnosing lung diseases.
摘要：
目的：据报道，血清克拉拉细胞分泌蛋白16（CC16）是肺损伤性疾病的潜在生物标志物。但是目前，没有其他方法更快，更准确,或更敏感的应用于临床实践中，除了ELISA。本研究旨在建立一种磁性纳米粒子化学发光免疫测定法（MNPs-CLIA），用于高灵敏地自动检测血清克拉拉细胞分泌蛋白16（CC16），并验证了其对肺部疾病的诊断性能。
方法：本研究包括CC16重组蛋白的表达，单克隆抗体(MAb)的制备与筛选,以及建筑，MNPs-CLIA方法的优化和分析评价。通过检测296份血清样本中CC16水平,探讨该方法的临床应用价值。
结果：MNPs-CLIA测定系统的线性范围为0.2-50ng/mL，检测限为0.037ng/mL。性能参数，如特异性，回收率,和精密度能满足体外诊断试剂的行业标准。该方法与目前临床使用的ELISA(R2=0.9962)结果一致,它还表现出令人满意的矽肺诊断功效，慢性阻塞性肺疾病(COPD),和肺结节病，曲线下面积（AUC）分别为0.9748、0.8428和0.9128。
结论：我们建立的MNPs-CLIA方法具有自动化的优点，高吞吐量,快速性,和简单，并可在临床推广应用。血清CC16的MNPs-CLIA检测对预测和诊断肺部疾病具有有效的诊断潜力。
方法：本研究包括CC16重组蛋白的表达，单克隆抗体(MAb)的制备与筛选,以及建筑，MNPs-CLIA方法的优化和分析评价。通过检测296份血清样本中CC16水平,探讨该方法的临床应用价值。
结果：MNPs-CLIA测定系统的线性范围为0.2-50ng/mL，检测限为0.037ng/mL。性能参数，如特异性，回收率,和精密度能满足体外诊断试剂的行业标准。该方法与目前临床使用的ELISA(R2=0.9962)结果一致,它还表现出令人满意的矽肺诊断功效，慢性阻塞性肺疾病(COPD),和肺结节病，曲线下面积（AUC）分别为0.9748、0.8428和0.9128。
结论：我们建立的MNPs-CLIA方法具有自动化的优点，高吞吐量,快速性,和简单，并可在临床推广应用。血清CC16的MNPs-CLIA检测对预测和诊断肺部疾病具有有效的诊断潜力。
