State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, Beijing 100071, China; The Fifth School of Clinical Medicine, Anhui Medical University, Hefei 230032, China; Department of Critical Care Medicine, The Fifth Medical Center of Chinese PLA General Hospital, Beijing 100071, China.
Department of Critical Care Medicine, The Fifth Medical Center of Chinese PLA General Hospital, Beijing 100071, China.
J Immunol Methods. 2023 Nov;522:113570. doi: 10.1016/j.jim.2023.113570. Epub 2023 Sep 27.
Human neutrophil lipocalin (HNL) is a marker of neutrophil activation and has a high efficacy in diagnosing bacterial infections. In this study, we applied the AlphaLISA technique to measure the serum level of HNL, evaluate HNL's efficacy in diagnosing septic shock, and identify any association between HNL level and septic patients' prognosis.
We collected 146 serum samples from the Fifth Medical Center of Chinese PLA General Hospital. HNL was measured by AlphaLISA and results were compared with commercial ELISA kits. We studied 78 patients admitted to the ICU with sepsis and data on their clinical and physiological characteristics were recorded. Blood levels of HNL, procalcitonin (PCT), high-sensitivity C-reactive protein (hs-CRP), and lactate were measured. A receiver operating characteristic (ROC) curve was used to evaluate the performance of each marker.
The AlphaLISA assay for serum HNL had a detection range from 1.5 ng/mL to 1000 ng/mL, with a detection limit of 1 ng/mL and a detection time of approximately 25 min. The AlphaLISA assay's results were in high agreement with ELISA results (R = 0.9413). HNL levels were analyzed in sepsis patients, and HNL was significantly higher in sepsis patients with shock compared to sepsis patients without shock (median 356.47 ng/mL vs 158.93 ng/mL, P < 0.0001) and in the 28-day non-survivor group compared to the 28-day survivor group (median 331.83 ng/mL vs 175.17 ng/mL, P < 0.0001). ROC curve analysis was performed for the biomarkers. In differentiating the diagnosis of septic shock from sepsis patients, HNL was the most effective marker (AUC = 0.857), followed by PCT (AUC = 0.754) and hs-CRP (AUC = 0.627). In predicting the prognosis of septic patients, lactate had the best effect (AUC = 0.805), followed by HNL (AUC = 0.784), PCT (AUC = 0.721), and hs-CRP (AUC = 0.583).
As an assessment tool, we found that our AlphaLISA had good consistency with an ELISA and had several other advantages, including requiring a shorter processing time and detecting a wider range of serum HNL concentrations. Monitoring serum HNL levels of patients admitted to the ICU might be useful in distinguishing sepsis patients who have septic shock from other sepsis patients, indicating its value in the prediction of sepsis patient prognosis.
人中性粒细胞明胶酶相关脂质运载蛋白（HNL）是一种中性粒细胞激活的标志物，在诊断细菌感染方面具有很高的功效。在这项研究中，我们应用 AlphaLISA 技术来测量血清 HNL 水平，评估 HNL 对诊断感染性休克的效果，并确定 HNL 水平与感染性休克患者预后之间的任何关联。
我们收集了来自中国人民解放军总医院第五医学中心的 146 份血清样本。通过 AlphaLISA 测量 HNL，结果与商业 ELISA 试剂盒进行比较。我们研究了 78 名入住 ICU 的患有败血症的患者，记录了他们的临床和生理特征数据。测量了 HNL、降钙素原（PCT）、高敏 C 反应蛋白（hs-CRP）和乳酸的血水平。使用受试者工作特征（ROC）曲线评估每个标志物的性能。
AlphaLISA 测定血清 HNL 的检测范围为 1.5ng/mL 至 1000ng/mL，检测限为 1ng/mL，检测时间约为 25 分钟。AlphaLISA 测定结果与 ELISA 结果高度一致（R=0.9413）。分析败血症患者的 HNL 水平，与无休克的败血症患者相比，休克的败血症患者的 HNL 水平显著升高（中位数 356.47ng/mL 比 158.93ng/mL，P<0.0001），与 28 天非幸存者组相比，28 天幸存者组的 HNL 水平也显著升高（中位数 331.83ng/mL 比 175.17ng/mL，P<0.0001）。对生物标志物进行 ROC 曲线分析。在区分感染性休克与败血症患者的诊断中，HNL 是最有效的标志物（AUC=0.857），其次是 PCT（AUC=0.754）和 hs-CRP（AUC=0.627）。在预测败血症患者的预后时，乳酸的效果最好（AUC=0.805），其次是 HNL（AUC=0.784）、PCT（AUC=0.721）和 hs-CRP（AUC=0.583）。
作为一种评估工具，我们发现我们的 AlphaLISA 与 ELISA 具有良好的一致性，并且具有其他几个优势，包括需要更短的处理时间和检测更宽范围的血清 HNL 浓度。监测入住 ICU 的患者的血清 HNL 水平可能有助于区分患有感染性休克的败血症患者和其他败血症患者，表明其在预测败血症患者预后方面的价值。
