Molecular Analysis of Pathogens in Otitis Media by PCR

PCR 法对中耳炎病原体进行分子分析

• 批准号: 7901732
• 负责人: Garth D. Ehrlich
• 金额: $ 25.42万
• 依托单位: ALLEGHENY-SINGER RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-14 至 2010-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7901732
• 关键词: Adenoidal structure; Adenoidectomy; Affect; Animals; Antibiotic Therapy; Antibiotics; Arts; Bacteria; Child; Chinchilla (genus); Chronic; Clinical; Congenic Strain; DNA Sequence Analysis; Data; Development; Disease; Economically Deprived Population; Gene Chips; Gene Expression; General anesthetic drugs; Genes; Genetic; Genetic Recombination; Genome; Genomics; Goals; Haemophilus influenzae; Health Personnel; Human; Image; Individual; Infection; Inflammatory; Investigation; Knock-out; Laboratories; Libraries; Light; Microbial Biofilms; Modeling; Molecular Analysis; Molecular Biology; Native Americans; Open Reading Frames; Oral; Organism; Otitis Media; Pathogenicity; Patients; Physiology; Population; Principal Investigator; Process; Proteomics; Recombinants; Reverse Transcriptase Polymerase Chain Reaction; Stress; Techniques; Testing; Time; Tissues; Treatment Failure; Tube; Tympanostomy; Virulence Factors; Visit; Woman; comparative; effective therapy; effusion; genome sequencing; in vivo; indexing; knockout gene; novel; pathogen; pathogenic bacteria; programs; protein expression; research study; success

DESCRIPTION (provided by applicant): Otitis media (OM) is the most common reason that an ill child visits a health care provider or undergoes a general anesthetic. OM is also the most common reason that a child receives an oral antibiotic and the over-treatment of patients with OM has been suspected of contributing to the development of antimicrobialresistant organisms. OM disproportionately affects socio-economically disadvantaged children, Native American children and is a factor that inhibits women from full participation in the workforce. The major focus of our laboratory for the past decade has been elucidating the path physiology of chronic OM with the ultimate goal of developing more effective treatments. During this time, we have shown that chronic OM is not a purely inflammatory process, but rather is a bacterial biofilm illness. The recognition that OM is a biofilm disease then led to a novel hypothesis: The Distributed Genome Hypothesis. This hypothesis states that there is a supra-genome for pathogenic bacteria and that each individual bacterium possesses only a subset of genes from the supra-genome. The supra-genome is a reservoir for panoply of contingency genes that collectively provides a significant survival benefit for the population-at-large. In this continuing application we specifically test the Distributed Genome Hypothesis in Haemophilus influenza (HI) with four Specific Aims: 1) Perform comparative genomic studies among clinical isolates of HI To characterize the extent of genomic plasticity; 2) Determine the extent of HI inter-isolate recombination during the infectious process; 3) Prepare transformation knockouts of HI and compare their survival time in vivo with wild-type congener strains; 4) Phenotypic characterization of HI biofilms. These Specific Aims will be accomplished by experiments using state-of-the-art high throughput genomic, molecular biology, imaging and modeling techniques, as well as investigations in children. Preliminary data generated from a micro array library composed of 10 clinical isolates of H. influenza demonstrated that H. influenza does indeed have a supra-genome that is twice the Size of a single bacterium. These findings shed light on many aspects of OM, including disease persistence in the fact of antibiotic treatment, and provide an explanation for the success of adenoidectomy in the management of OM.

描述（由申请人提供）：中耳炎 (OM) 是患病儿童去看医生或接受全身麻醉的最常见原因。 OM 也是儿童接受口服抗生素的最常见原因，并且 OM 患者的过度治疗被怀疑会导致抗菌药物耐药菌的产生。 OM 不成比例地影响社会经济弱势儿童、美洲原住民儿童，也是阻碍妇女充分参与劳动力的一个因素。 我们实验室过去十年的主要重点是阐明慢性 OM 的病理生理学，最终目标是开发更有效的治疗方法。在此期间，我们已经证明慢性 OM 并不是纯粹的炎症过程，而是一种细菌生物膜疾病。 OM 是一种生物膜疾病的认识引发了一个新的假说：分布式基因组假说。该假设指出，病原细菌存在一个超基因组，并且每个细菌仅拥有该超基因组基因的一个子集。超基因组是一整套偶然基因的储存库，这些基因共同为广大人口提供了显着的生存效益。在这个持续的应用中，我们专门测试了流感嗜血杆菌（HI）的分布式基因组假说，有四个具体目标：1）在HI临床分离株之间进行比较基因组研究，以表征基因组可塑性的程度； 2) 确定感染过程中HI分离株间重组的程度； 3) 制备HI的转化敲除株，并与野生型同源株进行体内存活时间比较； 4) HI 生物膜的表型特征。这些具体目标将通过使用最先进的高通量基因组、分子生物学、成像和建模技术的实验以及对儿童的研究来实现。由 10 种流感嗜血杆菌临床分离株组成的微阵列文库生成的初步数据表明，流感嗜血杆菌确实具有两倍于单个细菌大小的超基因组。这些发现揭示了 OM 的许多方面，包括抗生素治疗中疾病的持续存在，并为腺样体切除术在 OM 治疗中的成功提供了解释。