High through-put detection of fungal pathogens

真菌病原体的高通量检测

• 批准号: 6577634
• 负责人: JACK W FELL
• 金额: $ 37.99万
• 依托单位: UNIVERSITY OF MIAMI ROSENTEIL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6577634
• 关键词: Aspergillus; Candida; Cryptococcus; Fungi; biotechnology; clinical research; communicable disease diagnosis; cooperative study; diagnosis design /evaluation; flow cytometry; high throughput technology; mycosis; nucleic acid hybridization

DESCRIPTION (provided by applicant): Diagnoses of opportunistic fungal infections constitute an increasing clinical problem. Conventional diagnostic tests are time consuming and lack specificity and sensitivity for accurate and timely prognoses. The lack of precise and prompt diagnoses has undermined the health of patients and contributed to high mortality rates in immunocompromised individuals undergoing aggressive medical treatments. This condition has led to an increase in the emergence of new antifungal resistant strains and the need for rapid non-culture based methods for the detection of pathogens at species and strain levels. In cooperation with a unique team of researchers from government (USDA, Peoria, Ill), academia (University of Miami), two clinical laboratories (John Hopkins Medical Institute and Allogen Laboratories of the Cleveland Clinic) and industry (Luminex Corp, Austin, TX) we propose to develop a sensitive, fast and reliable high throughput DNA based method aimed at detection of such important fungal pathogens as species of Aspergillus, Candida, Cryptococcus and Trichosporon. The detection method will combine the specificity and reliability of nucleic acid hybridization analysis with the speed and sensitivity of LabMap, a new flow cytometric technique by Luminex Corp. The multiplexed assay analysis will consist of a combination of fluorescent microspheres covalently bound to species-specific fungal oligonucleotide probes, with the capability to simultaneously test as many as 100 different species-specific probes per tube/well at rates 0.47 min/well probes per minute. In the preliminary stages of this research, the system will be developed with extensive in-house databases and culture collections from USDA and UM and from clinical isolates from JHMI to produce a detection technique for cultured organisms. Validation of the assay with archived clinical specimens will be in conjunction with the Allogen Lab, which will lead to a detection system directly from clinical specimens. The fungal detection system will be expanded to include all known yeasts, including medical, foodborne and industrial species as well as other filamentous pathogens. We foresee that this multiplex testing fungal platform can be expanded to other microorganisms with important applications in areas of public health, bioterrorism, food safety and the environment. Resulting data will be disseminated via GenBank, publications in leading scientific journals and as an inexpensive commercial detection system.

描述（由申请人提供）：机会性真菌感染的诊断构成了越来越多的临床问题。常规的诊断测试是耗时的，缺乏对准确和及时预后的特异性和敏感性。缺乏精确和及时的诊断破坏了患者的健康状况，并导致接受积极医疗治疗的免疫功能低下的个体死亡率高。这种情况导致新的抗真菌抗真菌菌株的出现增加，并且需要基于非文化的快速方法来检测物种和应变水平的病原体。与政府（USDA，Peoria，Ill）的独特研究人员合作，学术界（迈阿密大学），两个临床实验室（约翰·霍普金斯医学院和克利夫兰诊所的Allogen实验室）和行业（Luminex Corp，TX，奥斯汀，德克萨斯州奥斯汀，德克萨斯州） ）我们建议开发一种敏感，快速和可靠的基于高吞吐DNA的方法，旨在检测诸如曲霉菌，念珠菌，加密摄氏和trichosporon等重要的真菌病原体。检测方法将将核酸杂交分析的特异性和可靠性与LabMap的速度和灵敏度结合在一起，Luminex Corp.一种新的流式细胞仪技术。多路复用测定分析将包括荧光微球共同结合到物种特异性的真实性真实性真实性真实性真菌的结合。寡核苷酸探针，其能力以每分钟0.47分钟/井探针的速率同时测试多达100个不同物种特异性探针。在这项研究的初步阶段，该系统将通过USDA和UM以及JHMI的临床分离株进行广泛的内部数据库和文化收集开发，以生成用于培养生物体的检测技术。用存档的临床标本对测定的验证将与Allogen Lab结合使用，该实验室将直接从临床标本中导致检测系统。真菌检测系统将扩展到包括所有已知的酵母菌，包括医疗，粮食生物和工业物种以及其他丝状病原体。我们预见，这种多重测试真菌平台可以扩展到其他微生物，在公共卫生，生物恐怖，食品安全和环境领域具有重要应用。所得数据将通过GenBank，领先的科学期刊的出版物以及廉价的商业检测系统传播。