DETECTING AND QUANTITATING UNCULTURABLE HUMAN PATHOGENS

检测和定量不可培养的人类病原体

• 批准号: 2422360
• 负责人: NORMAN G ANDERSON
• 金额: $ 10万
• 依托单位: LARGE SCALE BIOLOGY CORPORATION
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-07-01 至 1998-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2422360
• 关键词: biotechnology; communicable disease diagnosis; density gradient ultracentrifugation; diagnosis design /evaluation; fluorescent dye /probe; lasers; method development; particle counter; serology /serodiagnosis; virus characteristic; virus load

This proposal is to develop a rapid method for separating, identifying and counting small numbers of viral pathogens from tissues, blood, urine, water supplies, and natural waters, with emphasis on agents currently unculturable. In nature there are few particles of viral dimensions, which have both a narrow range of both sedimentation rates and isopycnic banding densities which are not viruses. Hence it is feasible to develop centrifugal systems to concentrate particles in the desired size range, and, using a new centrifuge tube under development, to concentrate and isopycnically band viral particles from several milliliters to a band having a volume of a cubic micron in a specially designed small-diameter density gradient. Using blue laser illumination the particles may be detected by light scattering or fluorescence using new fluorescent dyes which penetrate viruses and stain DNA or RNA. With these dyes, single virions have been detected. We believe the system is applicable to the search for previously undetected viruses, and to clinically detecting and quantitating viremias in HIV and hepatitis A, B, and C and other infections, and to the problem of routinely distinguishing viral from bacterial infections. In Phase I a prototype system will be evaluated using model non-pathogenic viruses. PROPOSED COMMERCIAL APPLICATION: For research purposes we estimate a world market of approximately 200 machines, and for clinical diagnosis a market of over 2,000 machines.

本研究旨在建立一种快速分离、鉴定和鉴定大肠杆菌的方法。 从组织，血液，尿液， 水供应和天然沃茨，重点是目前 没有教养 自然界中几乎没有病毒大小的粒子， 其沉积速率和等密度线的范围都很窄 不是病毒的带密度。 因此，发展 离心系统将颗粒浓缩在所需的尺寸范围内， 使用正在开发的新型离心管， 等密度带状病毒颗粒从几毫升到一条带 在特别设计的小直径中具有立方微米的体积， 密度梯度 使用蓝色激光照射，颗粒可以是 用新的荧光染料通过光散射或荧光检测 能穿透病毒并染色DNA或RNA 有了这些染料， 已经检测到病毒粒子。 我们相信该系统适用于 搜索以前未检测到的病毒，并在临床上检测和 定量HIV和甲型、B和丙型肝炎及其他 感染，以及常规区分病毒和 细菌感染 在第一阶段，将对原型系统进行评估 使用模型非致病性病毒。 建议的商业应用：出于研究目的，我们估计 全球市场约有200台机器，临床诊断和 超过2,000台机器的市场。