TB Diagnostics at the Point of Care

护理点结核病诊断

• 批准号: 8073653
• 负责人: Christopher Gerard Cooney
• 金额: $ 21.01万
• 依托单位: AKONNI BIOSYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8073653
• 关键词: African American; Asians; Bedside Testings; Biological Assay; British Columbia; Centers for Disease Control and Prevention (U.S.); Clinic; Code; Communities; Community Healthcare; Commuting; Cytolysis; DNA; Devices; Diagnostic; Disease; Drug Resistant Tuberculosis; Drug resistance; Elements; Gel; Genes; Genetic Polymorphism; Genomics; Genotype; Goals; Gold; Hand; Hawaiian population; Heating; Laboratories; Latino; Methodology; Methods; Mexico; Minority; Multidrug-Resistant Tuberculosis; Mutation; Mycobacterium tuberculosis; Native Americans; Nucleic Acids; Pacific Island Americans; Persons; Phase; Physicians; Population; Preparation; Prevalence; Printing; Promoter Regions; Reaction; Ribosomal RNA; Sampling; Sensitivity and Specificity; Specificity; Sputum; System; Technology; Testing; Translating; Tuberculosis; United States; Work; base; biochip; cost; follow-up; health disparity; instrument; instrumentation; killings; member; microbial; point of care; prototype; public health relevance; resistant strain; socioeconomics

DESCRIPTION (provided by applicant): Of all diseases, Tuberculosis (TB) represents one of, if not, the greatest health disparity between whites and minorities [1]. To be specific, for every TB-infected white person in the United States, there are an estimated 9 African-Americans, 8 Latinos, 6 Native Americans, 23 Asians, and 21 Native Hawaiian/Pacific Islanders with this disease [2]. Compounded with this disparity is the prevalence of drug-resistant mutations of TB, which have an associated 1000 polymorphisms that span 36 genes, two promoter regions, and one ribosomal RNA coding region [3]. Current methodologies, available primarily to affluent healthcare communities, utilize microbial cultures, which require sophisticated laboratories and weeks before a result can be determined. Difficulties for minorities in a low socioeconomic class to commute and/or follow up with their physicians can result in a lack of appropriate treatment. A low-cost simple and rapid point-of-care (POC) test could expand drug-resistant TB diagnostics to these minority communities. However, current technologies lack sensitivity, specificity, and/or multiplexing capacity. We, therefore, propose to develop a POC device that offers the sensitivity of culture methods, specificity of nucleic acid methods, and a broad coverage of mutations. To accomplish this, we will expand upon our existing MDR-TB PCR-Microarray Biochips. These biochips consist of printed gel-element microarrays that have been shown to amplify target with immobilized primers in the gel elements. Previous work showed that at least 60 independent reactions can simultaneously amplify 1000, and in some cases 100 genomic copies, without needing to split, and thus dilute, the sample. Our team includes the Laboratorios Medicos Especializados in Juarez, Mexico. Team members from this facility will initially evaluate our sample purification device for Mycobacterium tuberculosis (MTB), previously shown to be sucessful at the hands of the British Columbia Centre for Disease Control (BC-CDC). Additionally, the Juarez team will verify Akonni's MDR-TB PCR-Microarray Biochip. In parallel, Akonni will expand the multiplexing capacity of the drug-resistant TB arrays, develop a lysis method, and translate the MDR-TB assay to Akonni's POC prototype device. During Phase II, the genotyping capacity will be expanded further and the POC devices will be translated to the Juarez clinic. This proposed test is projected to be a $3 consumable, operated on a $5000 instrument. (PUBLIC HEALTH RELEVANCE STATEMENT): Of all diseases, Tuberculosis (TB) represents one of, if not, the greatest health disparity between whites and minorities. To be specific, for every TB-infected white person in the United States, there are an estimated 9 African-Americans, 8 Latinos, 6 Native Americans, 23 Asians, and 21 Native Hawaiian/Pacific Islanders with this disease. The proposed project is to develop a point-of-care device for identifying drug-resistant strains of Tuberculosis that can be widely disseminated to minority populations.

描述（由申请人提供）：在所有疾病中，结核病 (TB) 是白人和少数族裔之间最大的健康差距之一（如果不是的话）[1]。具体而言，在美国，每感染 TB 的白人中，估计就有 9 名非裔美国人、8 名拉丁裔、6 名美洲原住民、23 名亚洲人和 21 名夏威夷原住民/太平洋岛民患有这种疾病 [2]。与这种差异相结合的是结核病耐药突变的流行，这种突变具有 1000 个相关的多态性，涵盖 36 个基因、两个启动子区域和一个核糖体 RNA 编码区域 [3]。目前的方法主要适用于富裕的医疗保健社区，利用微生物培养，这需要复杂的实验室，并且需要数周才能确定结果。社会经济地位较低的少数群体在通勤和/或跟进医生方面存在困难，可能会导致缺乏适当的治疗。低成本、简单、快速的现场护理 (POC) 测试可以扩展 对这些少数民族社区进行耐药结核病诊断。然而，当前的技术缺乏敏感性、特异性和/或多路复用能力。 因此，我们建议开发一种 POC 设备，提供培养方法的敏感性、核酸方法的特异性以及广泛的突变覆盖范围。为了实现这一目标，我们将扩展现有的耐多药结核病 PCR 微阵列生物芯片。这些生物芯片由印刷的凝胶元件微阵列组成，已被证明可以通过凝胶元件中固定的引物来扩增靶标。之前的工作表明，至少 60 个独立反应可以同时扩增 1000 个（在某些情况下 100 个）基因组拷贝，而无需拆分样品，从而稀释样品。 我们的团队包括位于墨西哥华雷斯的 Laboratorios Medicos Especializados。该设施的团队成员将首先评估我们的结核分枝杆菌 (MTB) 样本纯化装置，该装置此前在不列颠哥伦比亚省疾病控制中心 (BC-CDC) 的使用中已被证明是成功的。此外，Juarez 团队还将验证 Akonni 的 MDR-TB PCR 微阵列生物芯片。与此同时，Akonni 将扩大耐药 TB 阵列的多重检测能力，开发裂解方法，并将 MDR-TB 检测转化为 Akonni 的 POC 原型设备。在第二阶段，基因分型能力将进一步扩大，POC设备将被转移到华雷斯诊所。这项提议的测试预计需要 3 美元的消耗品，在价值 5000 美元的仪器上运行。 （公共卫生相关声明）：在所有疾病中，结核病（TB）即使不是也是白人和少数族裔之间最大的健康差距之一。具体来说，在美国，每有一个感染结核病的白人，估计就有 9 名非洲裔美国人、8 名拉丁裔美国人、6 名美洲原住民、23 名亚洲人以及 21 名夏威夷原住民/太平洋岛民患有这种疾病。拟议的项目是开发一种用于识别结核病耐药菌株的即时护理设备，该设备可以广泛传播给少数群体。

项目成果

A bench-top automated workstation for nucleic acid isolation from clinical sample types.

用于从临床样本类型中分离核酸的台式自动化工作站。

• DOI: 10.1016/j.mimet.2018.03.021
• 发表时间: 2018
• 期刊: Journal of microbiological methods
• 影响因子: 2.2
• 作者: Thakore,Nitu;Garber,Steve;Bueno,Arial;Qu,Peter;Norville,Ryan;Villanueva,Michael;Chandler,DarrellP;Holmberg,Rebecca;Cooney,ChristopherG
• 通讯作者: Cooney,ChristopherG