Automated assay of plasma (1*3) -beta-D-glucan for early diagnosis of deep mycosis

血浆 (1*3)-β-D-葡聚糖自动检测用于早期诊断深部真菌病

基本信息

批准号：
06672300
负责人：
OBAYASHI Taminori
金额：
$ 1.28万
依托单位：
Jichi Medical School (JMS)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06672300/
关键词：
deep mycosis (1*3) -beta-D-glucan triple helix single strand automation horseshoe crab (1*3) -beta-D-glucan binding protein Factor G カブトガニ血球抽出液抗(1-3)-β-D-グルカン抗体 ELISA 三重鎖螺旋構造カイネティック・モード

项目摘要

Automatic assay of plasma (1*3) -beta-D-glucan with limulus reagent has been materialized with a new pretreating mixture which obviates the necessity of centrifugation. The mixture consists of KOH,KCL,and polybrene. KOH is to remove limulus-test interfering factors and enhance the sensitivity of the test by dissembling the triple helix of the glucan into single strands, which are a more potent activator of factor G.KCL is to keep the dissembled glucan in single strands, and polybrene is to prevent the formation of nonspecific turbidity. The adoption of kinetic mode has broadened the dynamic range up to 500 pg/ml. With a cutoff of 20 pg/ml, the sensitivity and the specificity of the test for invasive deep mycoses were 90% and 100%, respectively.Recently a new protein was isolated from horseshoe crab hemolymph that specifically binds to (1*3) -beta-D-glucan, thereby inhibiting the activation of factor G by the glucan. Using this new protein and anti- (1*3) -beta-D-glucan antibody, we developed an ELISA for (1*3) -beta-D-glucan determination. Plasma glucan concentrations in invasive deep mycoses showed a good correlation between the two methods, another evidence that the glucan is increased in blood in this conditon.

使用鲎试剂自动测定血浆（1*3）-β-D-葡聚糖已通过一种新的预处理混合物实现，从而消除了离心的必要性。该混合物由 KOH、KCL 和聚凝胺组成。 KOH是将葡聚糖的三螺旋分解成单链，去除鲎试验的干扰因素，提高测试的灵敏度，这是更有效的G因子激活剂。KCL是将分解的葡聚糖保持在单链，聚凝胺是为了防止非特异性浊度的形成。动力学模式的采用将动态范围扩大至 500 pg/ml。以 20 pg/ml 为截止值，该检测对侵袭性深部真菌病的敏感性和特异性分别为 90% 和 100%。最近从鲎血淋巴中分离出一种新蛋白质，该蛋白质特异性结合 (1*3) - β-D-葡聚糖，从而抑制葡聚糖对G因子的激活。使用这种新蛋白质和抗 (1*3) -β-D-葡聚糖抗体，我们开发了用于 (1*3) -β-D-葡聚糖测定的 ELISA。侵袭性深部真菌病中的血浆葡聚糖浓度显示两种方法之间具有良好的相关性，这是这种情况下血液中葡聚糖增加的另一个证据。