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种类是世界各地的主要病原体。它们是一个更大的所谓挑剔病原体群体的一部分，这些病原体很难研究，因为由于各种原因，它们很难培养。表型MicroArrayTM(PM)技术是一种有助于了解挑剔病原体的生理和代谢特性的工具。它使科学家能够在一次实验中扫描近2000种微生物细胞系的表型。这项技术现在可以用于对人类健康感兴趣的大多数需氧革兰氏阴性/阳性物种，70多份科学出版物和演示文稿展示了如何将其应用于广泛的科学调查。然而，一些挑剔的属目前不适用于PM分析。该STTR项目的一个主要目标是使PM技术适用于这些挑剔的需氧、微需氧和厌氧细菌种类，特别强调那些在人类疾病和/或生物防御方面的重要考虑因素。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、诺卡氏菌、军团菌)，嗜氧胃肠道病原体(螺杆菌、弯曲杆菌、弧菌、狼疮杆菌)，以及结肠和阴道的重要定殖菌(类杆菌、梭状芽孢杆菌、乳杆菌、大肠杆菌)。这些被选为最优先的项目，因为我们已经收到了40多名科学家的请求，他们希望在这些微生物的研究中利用PM技术。这种程度的科学兴趣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微生物识别系统的能力，该系统目前安装在大约一半的美国州立公共卫生实验室中，经常用于识别不太常见或不常见的微生物病原体。这些属中的大多数目前也没有包括在Biolog的识别数据库中。然而，随着这里提出的开发努力取得成功，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，我们在德克萨斯农工大学的合作者将专注于最重要的分枝杆菌属，并使用PM技术的敲除菌株来研究与该属的新陈代谢和耐药性有关的独特基因的功能。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有效地同时测试数百到数千种细胞表型的技术，直接造福于公众健康。它将应用于医院诊断工作、环境诊断工作、生物恐怖主义监测、新药开发、毒理学研究以及许多基础生物学研究领域。

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.