Phenotype MicroArray Analysis of Fastidious Pathogens

描述（申请人提供）：结核分枝杆菌和其他分枝杆菌属是世界各地的主要病原体。它们是一个更大的所谓挑剔的病原体群体的一部分，这些病原体很难研究，因为出于各种原因，它们很难培养。Phenotype MicroArrayTM（PM）技术是一种工具，可以帮助了解挑剔病原体的生理和代谢特性。它使科学家能够在一次实验中扫描近2,000种微生物细胞系的表型。这项技术现在可以用于人类健康中感兴趣的大多数需氧革兰氏阴性/阳性菌种，70多个科学出版物和演示文稿展示了它如何应用于广泛的科学研究。然而，一些挑剔的属目前不适合PM分析。该STTR项目的一个主要目标是使PM技术适应这些苛刻的好氧、微需氧和厌氧细菌物种，特别强调那些在人类疾病和/或生物防御方面重要的细菌。我们将重点关注的属包括肺部、皮肤和组织感染的病原体（分枝杆菌属、诺卡氏菌属、军团菌属）、微需氧胃肠病原体（螺杆菌属、弯曲杆菌属、弓形杆菌属、Wolinella）以及结肠和阴道的重要定植菌（拟杆菌属、梭菌属、乳杆菌属、埃希氏菌属）。这些被选为最高优先级，因为我们已经收到了40多位科学家的请求，他们希望在这些微生物的研究中利用PM技术。这种科学兴趣水平证明了我们提议开发的技术商业化的必要性和潜力。第二个目标是扩大Biolog微生物鉴定系统的能力，该系统目前安装在大约一半的美国州公共卫生实验室，经常用于鉴定不太常见或不寻常的微生物病原体。这些属中的大多数目前也没有包括在Biolog的鉴定数据库中。然而，随着这里提出的发展努力的成功，数据库将扩大到包括这些重要的挑剔物种。作为第三个目标，我们在德克萨斯A&M大学的合作者将专注于最重要的属-分枝杆菌，并使用PM技术的敲除菌株来研究该属代谢和耐药性所特有的基因的功能。如果成功，该项目的这一方面将有助于开发新的或更有效的抗分枝杆菌药物疗法。 该项目将通过提供一种同时有效检测数百至数千种细胞表型的技术，直接造福于公共卫生。它将应用于医院诊断工作、环境诊断工作、生物恐怖监测、新药开发、毒理学研究以及基础生物学研究的许多领域。

DESCRIPTION (provided by applicant): Mycobacterium tuberculosis and other Mycobacterium species are major pathogens around the world. They are part of a larger group of so-called fastidious pathogens that are difficult to study because, for a wide range of reasons, they are difficult to culture. Phenotype MicroArrayTM (PM) technology is a tool that can aid in understanding the physiological and metabolic properties of fastidious pathogens. It enables a scientist to scan nearly 2,000 phenotypes of a microbial cell line in a single experiment. This technology can now be used with most aerobic gram negative/positive species of interest in human health and more than 70 scientific publications and presentations demonstrate how it can be applied in a wide range of scientific investigations. However, a number of fastidious genera are currently not amenable to PM analysis. One principal goal for this STTR project is to adapt PM technology for these fastidious aerobic, microaerophilic, and anaerobic bacterial species, with special emphasis on those important in human disease and/or biodefense considerations. The genera we will focus on include agents of lung, cutaneous, and tissue infections (Mycobacterium, Nocardia, Legionella), microaerophilic gastro-intestinal pathogens (Helicobacter, Campylobacter, Arcobacter, Wolinella), and important colonizers of the colon and vagina (Bacteroides, Clostridium, Lactobacillus, Escherichia). These were selected as highest priority because we have received requests from more than 40 scientists seeking to utilize PM technology in their studies of these microorganisms. This level of scientific interest documents the need and potential for commercialization of the technology that we are proposing to develop. A second goal is to expand the capabilities of the Biolog Microbial Identification System, which is now installed in about half of US State Public Health Labs and is frequently used to identify less common or unusual microbial pathogens. Most of these genera are also not currently included in Biolog's Identification Database. However, with success in development efforts proposed here, the Database will be expanded to include these important fastidious species. As a third goal, our collaborator at Texas A&M University will focus on the most important genus, Mycobacterium, and use knockout strains with PM technology to study the function of genes that are unique to the metabolism and drug resistance of this genus. If successful, this aspect of the project will aid efforts toward the development of new or more effective anti-mycobacterial drug therapies. This project will directly benefit public health by providing a technology for efficient simultaneous testing of hundreds to thousands of cellular phenotypes. It will be applied in hospital diagnostic work, environmental diagnostic work, bioterrorism monitoring, new drug development, toxicology studies, and in many areas of basic biology research.