James E A, Schmeltzer K, Ligler F S
Naval Research Laboratory, Washington, DC 20375-5348, USA.
Appl Biochem Biotechnol. 1996 Sep;60(3):189-202. doi: 10.1007/BF02783583.
The lipopolysaccharide endotoxin is the most powerful immune stimulant known and a causative agent in the clinical syndrome known as sepsis. Sepsis is responsible for more than 100,000 deaths annually, in large part due to the lack of a rapid, reliable, and sensitive diagnostic technique. This study describes the detection of LPS from E. coli at concentrations as low as 10 ng/mL, in 30 s using an evanescent wave fiber-optic biosensor. Polymyxin B, covalently immobilized onto the surface of the fiber-optic probe, selectively bound fluorescently labeled LPS. Unlabeled LPS was detected in a competitive assay format using labeled LPS for signal generation. The competitive assay format worked in both buffer and plasma with similar sensitivities. This method can be used with other LPS capture molecules such as antibodies, lectins, or antibiotics, to simultaneously detect LPS and to determine the LPS serotype. The LPS assay using the fiber-optic biosensor is applicable to both clinical and environmental testing.
脂多糖内毒素是已知最强大的免疫刺激物,也是临床综合征脓毒症的致病因子。脓毒症每年导致超过10万例死亡,很大程度上是由于缺乏快速、可靠且灵敏的诊断技术。本研究描述了使用倏逝波光光纤生物传感器在30秒内检测低至10 ng/mL浓度的大肠杆菌脂多糖。共价固定在光纤探针表面的多粘菌素B选择性结合荧光标记的脂多糖。在竞争性检测模式中,使用标记的脂多糖产生信号来检测未标记的脂多糖。这种竞争性检测模式在缓冲液和血浆中均有效,且灵敏度相似。该方法可与其他脂多糖捕获分子(如抗体、凝集素或抗生素)一起使用,以同时检测脂多糖并确定脂多糖血清型。使用光纤生物传感器的脂多糖检测适用于临床和环境检测。