Martens A, Eppink G J, Woittiez A J, Eidhof H, de Leij L F
Laboratory of Clinical Chemistry, Twenteborg Hospital, Almelo, The Netherlands.
Crit Care Med. 1999 Mar;27(3):549-53. doi: 10.1097/00003246-199903000-00034.
To evaluate the performance of a newly developed assay to assess neutrophil function capacity. After optimization, the assay was performed on samples derived from patients with septic shock and compared with healthy controls and patients with a systemic viral infection.
Prospective evaluation of the performance of a new assay.
Medical intensive care unit, hospital laboratory.
Ten patients with septic shock, ten patients with infectious mononucleosis, and ten healthy controls.
We report an assay to assess neutrophil function capacity, in which CD10 membrane expression is measured by FACS before and after in vitro stimulation with Staphylococcus aureus bacteria. This assay evaluates the early activation state of circulating neutrophils and is shown to be of value in diagnosing a sepsis syndrome. First the assay was optimized. As an anticoagulant, sodium-citrate gave the best results. Blood samples must be kept on ice to reduce activation inside the siliconized tube and can be stored in this way for at least 8 hrs without affecting the test results. Kinetic studies showed a maximal expression of CD10 on neutrophils of healthy volunteers after 15 mins of stimulation with S. aureus bacteria. Second, the test was performed on samples derived from ten septic patients and ten patients with infectious mononucleosis. Septic patients had a significantly decreased CD10 expression capacity compared with healthy controls. Patients with infectious mononucleosis have a significantly higher CD10 expression capacity compared with septic patients, but in approximately one-half of them, the expression capacity was below the range found in controls.
These results indicate that in circulating neutrophils, the secretory vesicles have been mobilized completely in patients with septic shock. The assay proves to be of acceptable analytical quality and can be quickly and easily performed. Regarding clinical performance, this assay may be helpful in diagnosing septic shock.
评估一种新开发的用于评估中性粒细胞功能能力的检测方法。优化后,对脓毒性休克患者的样本进行该检测,并与健康对照者和系统性病毒感染患者进行比较。
对一种新检测方法的性能进行前瞻性评估。
医院重症监护病房、医院实验室。
10例脓毒性休克患者、10例传染性单核细胞增多症患者和10名健康对照者。
我们报告了一种评估中性粒细胞功能能力的检测方法,其中通过荧光激活细胞分选术(FACS)在体外用金黄色葡萄球菌刺激前后测量CD10膜表达。该检测方法评估循环中性粒细胞的早期激活状态,并且在诊断脓毒症综合征方面具有价值。首先对该检测方法进行了优化。作为抗凝剂,柠檬酸钠效果最佳。血样必须保存在冰上以减少硅化管内的激活,并且可以以这种方式保存至少8小时而不影响检测结果。动力学研究表明,健康志愿者的中性粒细胞在用金黄色葡萄球菌刺激15分钟后CD10表达达到最大值。其次,对10例脓毒症患者和10例传染性单核细胞增多症患者的样本进行了检测。脓毒症患者与健康对照者相比,CD10表达能力显著降低。传染性单核细胞增多症患者与脓毒症患者相比,CD10表达能力显著更高,但其中约一半患者的表达能力低于对照者的范围。
这些结果表明,在循环中性粒细胞中,脓毒性休克患者的分泌囊泡已被完全动员。该检测方法证明具有可接受的分析质量,并且可以快速、轻松地进行。关于临床性能,该检测方法可能有助于诊断脓毒性休克。