Rapid and Enclosed System for the Identification of Multi-Drug Resistant TB
用于识别多重耐药结核病的快速封闭系统
基本信息
- 批准号:7394908
- 负责人:
- 金额:$ 14.52万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2008
- 资助国家:美国
- 起止时间:2008-05-15 至 2010-07-31
- 项目状态:已结题
- 来源:
- 关键词:Academic Medical CentersAcquired Immunodeficiency SyndromeAfricaAgreementAirAntibioticsAntitubercular AgentsAreaArtsBacillus (bacterium)Biological AssayBlood capillariesBuffersCaliberCaringCause of DeathCessation of lifeCharacteristicsChemical EngineeringChinaClinicClinicalCollaborationsCollectionCommunicable DiseasesComplexConditionCountryCoupledDataData DisplayDepositionDetectionDeveloped CountriesDeveloping CountriesDevelopmentDevicesDiagnosisDiagnosticDiagnostic ProcedureDiagnostic ServicesDiagnostic testsDifferential DiagnosisDiffusionDimensionsDiseaseDrug Resistant TuberculosisDrug resistanceDrug resistance in tuberculosisDrug-sensitiveEncephalitisEngineeringEquipmentEvaluationFluorescenceFoundationsGlassGoalsGovernmentHIVHealth ProfessionalHealthcareHealthy People 2010HourHousingHumanImmune systemImmunologic Deficiency SyndromesIn VitroIncidenceIncomeIndividualInfectionInfectious Diseases ResearchInvestmentsKazakhstanLaboratoriesLateralLeadLengthLicensingLifeLiquid substanceMalariaMarketingMedicalMedicineMethodsMicrobeMicrobiologyMicrofluidicsMissionMolecularMolecular TargetMonitorMulti-Drug ResistanceMultidrug-Resistant TuberculosisMutationMycobacterium tuberculosisNatureNucleic AcidsNumbersOligonucleotidesPathologyPatientsPersonal SatisfactionPersonsPharmaceutical PreparationsPhasePliabilityPolymerase Chain ReactionPolymersPopulationPredispositionPreparationProceduresProcessProductionPublic HealthPumpPurposeRangeRateReactionReagentRecording of previous eventsReportingResearch PersonnelResistanceRespiratory Tract InfectionsRifampinRiskSamplingScientistScreening procedureSensitivity and SpecificityServicesSolidSolutionsSpecimenSputumStreamSurfaceSurveysSystemTechniquesTechnologyTechnology TransferTest ResultTestingTexasTimeTubeTuberculosisUSSRUnited StatesUniversitiesUniversity HospitalsViralWeekWorkWorld Health Organizationassay developmentbasecapillarycommercializationconceptcostdaydesigndisorder controldrug standardevaluation/testingexperiencefallsfoodborneinnovationinstrumentationinterestisoniazidkillingsmultidisciplinarynovel diagnosticsnucleic acid detectionpandemic diseasepathogenpreventprofessorrapid diagnosisreaction rateresearch and developmentstatisticssuccesstooltuberculosis drugstuberculosis treatment
项目摘要
DESCRIPTION (provided by applicant): Tuberculosis (TB) caused by Mycobacterium tuberculosis is the seventh leading cause of death worldwide, second only to human immunodeficiency virus (HIV) and acquired immunodeficiency syndrome (AIDS) among infectious diseases. In 2004, an estimated 8.9 million people developed TB and 1.7 million died. This situation is compounded by the HIV pandemic with almost 13 million people currently co-infected with HIV and TB, and the emergence of multidrug resistant TB (MDR-TB). In the Unites States, there are about 15,000 new cases of TB reported each year, and approximately 10% of them have resistance to drugs. Rapid diagnosis of TB infections and identification of drug resistance is key to disease control and treatment. In order to accomplish the Healthy People 2010 goal of reducing the time required for the laboratory confirmation of the diagnosis of tuberculosis to 48 hours, rapid tests to detect Mycobacterium tuberculosis or its products are needed. In addition, rapid tests that can reduce the turnaround-time for detection of drug- resistance are needed as well. It is clear that no test is yet available that meets target specifications, and new methods that can overcome limitations and respond to the challenges posed will be well received. Our proposed work will employ state-of-the-art technology to provide differential diagnostics to identify Mycobacterium tuberculosis infection and determine if it has mutations that cause resistances to the most commonly used antibiotics for TB. The goal is to achieve sample-to-result testing in less than 90 minutes. After sample preparation using automated nucleic acid (NA) extraction instrumentation, the NA template will be drawn into an in-line ultra-fast PCR reaction where a highly multiplexed amplification assay (covering 25 mutation/targets) will take place. This powerful multiplex amplification technology from our collaborator overcomes the obstacles of traditional multiplex PCR assay development. It allows for specific and sensitive amplification of multiple targets in a single reaction. The PCR products, mixed with hybridization buffer, will then be transported downstream in a closed format to perform an extremely simple microfluidic based multiplexed detection. The PCR product will be passed over a linear glass capillary substrate with spatially addressable capture zones that are modified with different oligonucleotide capture probes. The smaller diameter linear glass substrate will be housed in a slightly larger diameter polymeric tubing to provide an "array-in-a tube" conditions with microfluidic characteristics for rapid and sensitive fluorescence detection utilizing an extremely simple and inexpensive setup. In Phase I, a breadboard system will be developed and the technology feasibilities will be demonstrated with the nucleic acid templates provided by collaborators. Phase I results will enable the subsequent design and development of an automated clinical diagnostic assay that will fulfill a crucial need in TB care, as well as the detection of other diseases.
Tuberculosis (TB), which claims nearly 2 million lives each year, is the seventh leading cause of death worldwide, second only to HIV/AIDS among infectious diseases. Making the condition worse, the emergence of multidrug resistant TB (MDR-TB) makes the treatment more difficult and costly. In the Unites States, there are about 15,000 new cases of TB reported each year, and approximately 10% of them have resistance to drugs. A quick and accurate diagnostics tool for TB and MDR-TB is key to disease control and treatment and could save up to 625,000 of those lives each year (Nature S1, 46-57; 2006).
描述(申请人提供):由结核分枝杆菌引起的结核病(TB)是全球第七大致死原因,在传染病中仅次于人类免疫缺陷病毒(HIV)和获得性免疫缺陷综合征(AIDS)。2004年,估计有890万人患结核病,170万人死亡。艾滋病毒大流行使这种情况更加复杂,目前有近1300万人同时感染艾滋病毒和结核病,并出现了耐多药结核病。在美国,每年约有15,000例结核病新发病例,其中约10%对药物具有耐药性。结核病感染的快速诊断和耐药性鉴定是疾病控制和治疗的关键。为了实现《2010年健康人民》关于将实验室确认结核病诊断所需时间缩短至48小时的目标,需要快速检测结核分枝杆菌或其产物。此外,还需要能够缩短检测耐药性的周转时间的快速检测。很明显,目前还没有达到目标规格的检测方法,能够克服局限性和应对挑战的新方法将受到欢迎。我们拟议的工作将采用最先进的技术来提供鉴别诊断,以识别结核分枝杆菌感染,并确定它是否具有导致对结核病最常用抗生素耐药性的突变。我们的目标是在90分钟内完成从样品到结果的测试。在使用自动核酸(NA)提取仪器制备样品后,NA模板将被吸入在线超快速PCR反应中,其中将进行高度多重扩增测定(涵盖25个突变/靶标)。我们合作者的这种强大的多重扩增技术克服了传统多重PCR检测开发的障碍。它允许在单个反应中特异性和灵敏地扩增多个靶标。PCR产物与杂交缓冲液混合,然后以封闭形式向下游运输,以进行极其简单的基于微流体的多重检测。PCR产物将通过线性玻璃毛细管基底,该基底具有用不同寡核苷酸捕获探针修饰的空间可寻址捕获区。较小直径的线性玻璃基板将被容纳在直径稍大的聚合物管中,以提供具有微流体特性的“管中阵列”条件,用于利用极其简单和廉价的设置进行快速和灵敏的荧光检测。在第一阶段,将开发一个试验板系统,并利用合作者提供的核酸模板演示技术特性。第一阶段的结果将使随后的设计和开发的自动化临床诊断检测,将满足结核病护理的关键需求,以及其他疾病的检测。
结核病每年夺去近200万人的生命,是全世界第七大死亡原因,在传染病中仅次于艾滋病毒/艾滋病。使病情恶化的是,耐多药结核病(MDR-TB)的出现使治疗更加困难和昂贵。在美国,每年约有15,000例结核病新发病例,其中约10%对药物具有耐药性。结核病和耐多药结核病的快速准确诊断工具是疾病控制和治疗的关键,每年可以挽救多达625,000人的生命(Nature S1,46-57; 2006)。
项目成果
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SEASON S-S WONG其他文献
SEASON S-S WONG的其他文献
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