Optimized Growth Media to Detect Intracellular Bacteria Implicated in Chronic Dis
优化生长培养基以检测与慢性疾病有关的细胞内细菌
基本信息
- 批准号:8125369
- 负责人:
- 金额:$ 18.89万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2011
- 资助国家:美国
- 起止时间:2011-02-01 至 2012-01-31
- 项目状态:已结题
- 来源:
- 关键词:AddressAgarAlphaproteobacteriaAmericanAminoglycosidesAnimalsAntibioticsAntibodiesAnticoagulantsAntifungal AgentsAntineutrophil Cytoplasmic AntibodiesArthritisArthropod VectorsArthropodsAttentionAutoimmune ResponsesBacteriaBacterial InfectionsBartonellaBartonella InfectionsBartonella henselaeBartonella quintanaBiological AssayBiteBloodBlood donorBlood specimenCarbon DioxideCase SeriesCell DensityCenters for Disease Control and Prevention (U.S.)ChronicChronic DiseaseChronically IllClinicalClinical TrialsCoagulation ProcessCulture MediaCutaneousDNADataDetectionDevelopmentDiagnosisDiagnosticDiagnostics ResearchDiseaseDrug FormulationsEarly DiagnosisEndocarditisEpistaxisExpenditureFailureFleasFreezingFutureGalaxyGasesGoalsGranulomatousGrowthHeadHealthHealthcareHumanImmuneImmune systemImmunocompetentImpaired cognitionIncidenceIndividualInfectionInflammatoryLaboratoriesLeadLegal patentLiceLicensingLinkLiquid substanceLiteratureManufacturer NameMeasuresMedicalMedical ResearchMedicineMetabolismMethodsMicrobiologyMolecularMyocarditisNeedlesNeurologicNeurologic DysfunctionsNorth CarolinaOrgan failurePatientsPeliosis HepatisPhasePolyarthritidesPowder dose formPreventionProcessPropertyProtocols documentationRecoveryRegimenReportingResearchResearch DesignResearch PersonnelRickettsialesSalesSamplingSampling ErrorsSand FliesSavingsSpecimenSterilitySystemTechnologyTemperatureTestingTicksTimeUnited StatesUniversitiesVariantVasculitisVentVeterinary MedicineVial deviceVirulence FactorsWorkantimicrobialaxenic culturebasecell injurycollegecommercializationcommunicable disease diagnosisdesignexpectationexperienceflyimmunosuppressedimprovedinnovationinstrumentinterfacialmeetingsmembermicrobialnervous system disordernew growthnovelnovel diagnosticspathogenplanetary Atmosphereproduct developmentresearch studyresponsescale upsealtransmission processvector
项目摘要
DESCRIPTION (provided by applicant): The CDC reports that over 40 million Americans suffer from chronic disease, the majority of whom report multiple diagnoses of chronic disorders. The causes of many of these chronic diseases are not well understood and doctors struggle to effectively diagnose and treat these disorders. In recent years, attention has turned towards the possibility that many chronic disorders, especially those involving autoimmune response, may be caused by highly fastidious, immune-evasive pathogens. Bacteria of the genus Bartonella (a member of the Rickettsiales order) have been linked in the medical case series literature to a wide spectrum of diseases in human and veterinary medicine, including polyarthritis, cutaneous vasculitis, endocarditis, myocarditis, epistaxis, peliosis hepatis, granulomatous inflammatory disease, and, most recently, neurological dysfunction and arthritis. Traditional methods for detecting bacterial infections in patient samples have proven to be largely ineffective for the accurate diagnosis of Bartonellosis. Researchers at the North Carolina State University College of Veterinary Medicine have developed a highly sensitive Bartonella testing platform which combines the pre- enrichment culture of blood in a novel liquid media followed by a Bartonella PCR assay using genus level primers and sequence id of positive results. This innovative Bartonella testing platform was developed to facilitate the detection of Bartonella infections in immuno-competent patients where this highly fastidious bacteria infects at such low levels that sampling error at the point of testing introduces extremely high false negative rates for direct PCR and that the host immune system has little opportunity to develop antibodies against this bacteria. The cornerstone of this molecular diagnostic platform is a patented pre-enrichment medium called Bartonella Alpha Proteobacteria Growth Medium (BAPGM). Galaxy Diagnostics holds an exclusive license on BAPGM. Preliminary data support the usefulness of the BAPGM microbiological approach in human medicine. This novel Bartonella testing platform has resulted in the detection of single and polymicrobial Bartonella infections in blood samples obtained from both chronically-ill and healthy humans and animals. Perhaps more importantly research using the BAPGM-PCR testing platform have revealed important clinical associations between Bartonella infection and serious chronic disease. Our expectation is that the development of this testing platform will lead to medical breakthroughs in the diagnosis and treatment of chronic disease and will provide extensive savings in healthcare expenditures through early detection and prevention of debilitating chronic conditions. Of particular importance is research regarding modes of transmission and the identification as a cause or contributing factor in specific chronic disorders, like arthritis, neurological disease, organ failure and other hard- to-diagnose chronic conditions. As Bartonella spp. are transmitted by a large number of arthropod vectors, including fleas, lice, sand flies, ticks, biting flies and potentially others, determining the appropriate bacteria to screen for may be complicated by regional variations in the transmission of various Bartonella species to the blood donor public. Incidence of different Bartonella species by regions due to the existence of chronic carriers and/or vectors should be addressed in the future in order to establish the actual occurrence of this genus (and each of its species) in the US. The overall goal of Phase I will be to develop a BAPGM Axenic Culture Bottle formulation to be used for immediate inoculation or for transport of microbiological specimens. Specimens will be then transported to Galaxy Diagnostics for Bartonella testing using our current culture plus molecular testing protocol; Aim 1 develop and validate a new formulation in the standard BAPGM platform culture medium to fit the requirements of the BAPGM Axenic Culture Bottle design proposed; Aim 2 will test the feasibility that reasonable storage conditions on the new formulations can be achieved. This will be accomplished by examining growth sensitivity testing as performed previously after storage at room temperature, 4-8oC and -20oC. Phase II will focus on the product development and regulatory milestones required to develop test kits including the BAPGM Axenic Culture Bottle which will allow worldwide sale and distribution. Phase Ii research would also address the potential changes in the BAPGM Axenic Culture Bottle head-space gas composition required to maximize bacteria recovery and growth and that might lead to an optimization of the time required for the enrichment culture step in the testing platform.
描述(由申请人提供):CDC报告说,超过4000万美国人患有慢性疾病,其中大多数人报告慢性疾病的多重诊断。许多这些慢性疾病的原因还没有得到很好的理解,医生努力有效地诊断和治疗这些疾病。近年来,人们的注意力转向了许多慢性疾病,特别是涉及自身免疫反应的疾病,可能是由高度挑剔的免疫逃避性病原体引起的。在医学病例系列文献中,巴尔通体属的细菌(立克次体目的一个成员)与人类和兽医学中的广泛疾病有关,包括多关节炎、皮肤血管炎、心内膜炎、心肌炎、鼻衄、肝炎性紫癜、肉芽肿性炎性疾病,以及最近的神经功能障碍和关节炎。用于检测患者样本中细菌感染的传统方法已被证明对于准确诊断巴尔通体病基本无效。北卡罗来纳州州立大学兽医学院的研究人员开发了一种高灵敏度的巴尔通体测试平台,该平台将血液在新型液体培养基中的预富集培养物与随后使用属级引物和阳性结果的序列ID的巴尔通体PCR测定相结合。开发这种创新的巴尔通体检测平台是为了便于检测免疫功能正常的患者中的巴尔通体感染,其中这种高度挑剔的细菌以如此低的水平感染,以至于在检测时的采样误差为直接PCR引入了极高的假阴性率,并且宿主免疫系统几乎没有机会产生针对这种细菌的抗体。该分子诊断平台的基石是一种获得专利的预富集培养基,称为Bartonella Alpha Proteobacteria Growth Medium(BAPGM)。Galaxy Diagnostics拥有BAPGM的独家许可证。初步数据支持BAPGM微生物学方法在人类医学中的有用性。这种新的巴尔通体检测平台已导致从慢性病患者和健康人和动物获得的血液样本中检测到单一和多微生物巴尔通体感染。也许更重要的是,使用BAPGM-PCR检测平台的研究揭示了巴尔通体感染与严重慢性疾病之间的重要临床关联。我们的期望是,该检测平台的开发将导致慢性病诊断和治疗方面的医学突破,并将通过早期检测和预防使人衰弱的慢性病来节省大量的医疗支出。特别重要的是研究传播方式,并确定具体慢性疾病的原因或促成因素,如关节炎、神经系统疾病、器官衰竭和其他难以诊断的慢性病。巴尔通体属(Bartonella spp.)由于巴尔通体是由大量节肢动物传播的,包括跳蚤、虱子、白蛉、蜱、叮咬蝇和潜在的其他媒介,因此确定筛选的适当细菌可能会因各种巴尔通体物种传播给献血者公众的区域差异而变得复杂。由于存在慢性携带者和/或媒介,未来应解决不同地区巴尔通体属种的发病率,以确定该属(及其每个种)在美国的实际发生率。第I阶段的总体目标是开发用于立即接种或微生物标本运输的BAPGM无菌培养瓶配方。然后将样本运输至Galaxy Diagnostics,使用我们当前的培养加分子检测方案进行巴尔通体检测;目标1开发并验证标准BAPGM平台培养基中的新制剂,以符合拟定的BAPGM无菌培养瓶设计要求;目标2将检测新制剂可达到合理储存条件的可行性。这将通过在室温、4- 8 ℃和-20 ℃下储存后检查先前进行的生长敏感性试验来完成。第II阶段将重点关注开发检测试剂盒所需的产品开发和监管里程碑,包括允许全球销售和分销的BAPGM无菌培养瓶。第II阶段研究还将解决BAPGM无菌培养瓶顶部空间气体组成的潜在变化,以最大限度地提高细菌回收和生长,并可能导致测试平台中富集培养步骤所需时间的优化。
项目成果
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