Development of Indigenous Bio-sensing Methodology for Rapid and LowCost Endotoxin Detection System

B. Bal, Armstrong Pb, D. Ap
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引用次数: 7

Abstract

Endotoxin is a signature molecule of gram-negative bacteria and is clinically significant as the agent of gramnegative sepsis, a disease condition with high mortality. This study describes an enzyme-substrate reaction using the distilled water lysate of the granular hemocytes (amebocytes) of the mangrove horseshoe crab (Tachypleus gigas), a native of the Bay of Bengal, and a chromogenic peptide which results in the production of yellow spectroscopically monitored product in the presence of endotoxin. The assay is complete within 30 min and shows a lower detection limit of 0.2 EU/mL. The novelty of our approach is the use of mangrove horseshoe crabs are the source of amebocyte lysate instead of the American horseshoe crab (Limuluspoly phemus).The anticipated methodology offers advantages for the South Asian market that include a low cost due to use of indigenous reagents. This method shows exquisite sensitivity and provides a rapid assessment of LPS concentrations. We assume that the commercial development of our assay will reduce the cost of a single chromogenic endotoxin detection test as compared to the cost of present imported kits presently available in Asian and Indian market.
快速低成本内毒素检测系统的本土生物传感方法开发
内毒素是革兰氏阴性菌的一种特征分子,是革兰氏阴性脓毒症(一种高死亡率的疾病)的重要致病因子。本研究描述了一种酶-底物反应,使用孟加拉湾本地的红树林马蹄蟹(Tachypleus gigas)的颗粒血细胞(变形细胞)的蒸馏水裂解物和显色肽,在内毒素存在的情况下产生黄色光谱监测产品。该分析在30分钟内完成,并显示0.2 EU/mL的较低检测限。我们方法的新颖之处在于用红树林马蹄蟹代替美洲马蹄蟹(limuspoly phemus)作为鲎试剂的来源。预期的方法为南亚市场提供了优势,包括由于使用本土试剂而成本较低。该方法灵敏度高,可快速测定LPS浓度。我们认为,与目前在亚洲和印度市场上可用的进口试剂盒相比,我们的检测方法的商业发展将降低单次显色内毒素检测测试的成本。
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