Horváth-Szalai Zoltán, Kustán Péter, Szirmay Balázs, Lakatos Ágnes, Christensen Per H, Huber Tamás, Bugyi Beáta, Mühl Diána, Ludány Andrea, Miseta Attila, Kovács Gábor L, Kőszegi Tamás
Department of Laboratory Medicine, University of Pécs, Pécs, Hungary.
Department of Anaesthesiology and Intensive Therapy, University of Pécs, Pécs, Hungary.
J Clin Lab Anal. 2018 Mar;32(3). doi: 10.1002/jcla.22321. Epub 2017 Sep 5.
Studies showing the potential predictive value of the actin-binding protein gelsolin, in critically ill patients are scarce. Moreover, even up to now a rapid automated measurement of gelsolin has still remained a challenge. Therefore, we developed and validated an automated serum gelsolin immune turbidimetric assay for possible clinical use.
Validation of serum gelsolin assay was performed on a Cobas 8000/c502 analyzer (Roche) according to the second edition of Eurachem guidelines. Furthermore, we also studied the diagnostic value of serum gelsolin in sepsis when investigating sera of septic (n = 25), systemic inflammatory response syndrome (SIRS; n = 8) and control patients (n = 14). We compared our previously published Western blot data with those of the new turbidimetric assay.
The sample volume was 7 μL and the assay time was 10 minutes. The detection limit was 0.72 mg/L, intra- and inter-assay imprecision remained in most cases less than 5% expressed as CV. Recovery was found to be 84.56%-93.52% and linearity study gave an appropriate correlation coefficient by linear regression analysis (r = .998). Septic patients exhibited lower (P = .015) first-day serum gelsolin levels than SIRS patients, which confirmed our previous Western blot results. The determined cut-off point for serum gelsolin was 14.05 mg/L (sensitivity: 75%; specificity: 60%) when investigating its diagnostic value in sepsis.
Based on the results, our immune turbidimetric measurement offers a rapid and accurate quantitation of gelsolin in human serum samples. Serum gelsolin seems a promising additional diagnostic marker of sepsis which has to be further investigated.
关于肌动蛋白结合蛋白凝溶胶蛋白在危重症患者中潜在预测价值的研究较少。此外，直至目前，快速自动化检测凝溶胶蛋白仍然是一项挑战。因此，我们开发并验证了一种可用于临床的自动化血清凝溶胶蛋白免疫比浊测定法。
根据Eurachem指南第二版，在Cobas 8000/c502分析仪（罗氏公司）上对血清凝溶胶蛋白测定法进行验证。此外，在研究脓毒症患者（n = 25）、全身炎症反应综合征（SIRS；n = 8）患者及对照患者（n = 14）的血清时，我们还研究了血清凝溶胶蛋白在脓毒症中的诊断价值。我们将之前发表的蛋白质印迹数据与新的比浊测定法数据进行了比较。
样本体积为7μL，测定时间为10分钟。检测限为0.72mg/L，批内和批间不精密度在大多数情况下以CV表示小于5%。回收率为84.56% - 93.52%，线性研究通过线性回归分析得出了合适的相关系数（r = 0.998）。脓毒症患者第一天的血清凝溶胶蛋白水平低于SIRS患者（P = 0.015），这证实了我们之前的蛋白质印迹结果。在研究血清凝溶胶蛋白在脓毒症中的诊断价值时，其确定的截断点为14.05mg/L（敏感性：75%；特异性：60%）。
基于这些结果，我们的免疫比浊测定法能够快速、准确地定量检测人血清样本中的凝溶胶蛋白。血清凝溶胶蛋白似乎是脓毒症一个有前景的辅助诊断标志物，有待进一步研究。
