Yokota M, Kambayashi J, Tsujinaka T, Sakon M, Mori T, Tsuchiya M, Oishi H, Matsuura S
Second Department of Surgery, Osaka University Medical School, Japan.
Jpn J Surg. 1990 Sep;20(5):559-66. doi: 10.1007/BF02471013.
In order to correctly diagnose and treat severe postoperative infections, it may be critical to detect and differentiate between endotoxin derived from Gram-negative bacteria and/or beta-glucan derived from fungi. In addition to the chromogenic assay, the turbidimetric kinetic assay has been performed for the quantification of endotoxin in plasma using Limulus amebocyte lysate as previously reported. However, it is also known that beta-glucan triggers the coagulation of Limulus amebocyte lysate. In the present study, the differentiation of beta-glucan from endotoxin and its clinical application were studied. Endotoxin was able to be inactivated in plasma using one-tenth dilution by 10 per cent ethanol or distilled water, followed by heating at 100 degrees C for 120 min, without affecting the activity of coexisting beta-glucan. The treated sample was then subjected to the turbidimetric kinetic assay using Toxinometer ET-201. Using this method, as little as 30 pg/ml of beta-glucan in the plasma may be assayed separately, with the amount of circulating beta-glucan in the plasma of normal subjects being less than 50 pg/ml. On the other hand, in patients with a fungal infection, the amount of beta-glucan in their plasma was elevated significantly. Clinically, beta-glucanemia may often occur in severe postoperative infection even if fungi are not detected.
为了正确诊断和治疗严重的术后感染,检测并区分革兰氏阴性菌衍生的内毒素和/或真菌衍生的β-葡聚糖可能至关重要。除了显色测定法外,如先前报道的那样,还使用鲎试剂进行了比浊动力学测定以定量血浆中的内毒素。然而,众所周知,β-葡聚糖会引发鲎试剂的凝固。在本研究中,对β-葡聚糖与内毒素的区分及其临床应用进行了研究。使用10%乙醇或蒸馏水进行十分之一稀释,然后在100℃加热120分钟,可使血浆中的内毒素失活,且不影响共存的β-葡聚糖的活性。然后使用Toxinometer ET-201对处理后的样品进行比浊动力学测定。使用该方法,血浆中低至30 pg/ml的β-葡聚糖可单独检测,正常受试者血浆中循环β-葡聚糖的量小于50 pg/ml。另一方面,在真菌感染患者中,其血浆中β-葡聚糖的量显著升高。临床上,即使未检测到真菌,严重术后感染中也常出现β-葡聚糖血症。