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）代表了白人和少数民族之间最大的健康差距之一。具体来说，在美国，每一个结核病感染的白色人中，估计有9个非洲裔美国人，8个拉丁美洲人，6个美洲原住民，23个亚洲人和21个夏威夷/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，这些突变具有相关的1000个多态性，跨越36个基因、两个启动子区和一个核糖体RNA编码区[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和数周才能确定结果。社会经济地位低下的少数群体难以与医生沟通和/或跟进，这可能导致缺乏适当的治疗。一种低成本的简单快速的即时检测（POC）可以扩展到 为这些少数民族社区提供耐药结核病诊断。然而，目前的技术缺乏灵敏度、特异性和/或多路复用能力。 因此，我们建议开发一种POC装置，该装置提供培养方法的灵敏度、核酸方法的特异性和突变的广泛覆盖。为了实现这一目标，我们将扩展现有的耐多药结核PCR微阵列生物芯片。这些生物芯片由打印的凝胶元件微阵列组成，已经显示出用凝胶元件中的固定引物扩增靶。以前的工作表明，至少60个独立的反应可以同时扩增1000个，在某些情况下100个基因组拷贝，而不需要分裂，从而稀释样品。 我们的团队包括墨西哥华雷斯的医疗专家。该机构的团队成员将首先评估我们的结核杆菌（MTB）样本纯化设备，该设备此前已被证明在不列颠哥伦比亚省疾病控制中心（BC-CDC）的支持下取得了成功。此外，华雷斯团队将验证Akonni的MDR-TB PCR微阵列生物芯片。与此同时，Akonni将扩大耐药结核病阵列的多路复用能力，开发一种裂解方法，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器械将转移到华雷斯诊所。该拟议测试预计为3美元的耗材，在5000美元的仪器上操作。 在所有疾病中，结核病是白人和少数民族之间最大的健康差距之一。具体来说，在美国，每一个结核病感染的白色人中，估计有9个非洲裔美国人，8个拉丁美洲人，6个美洲原住民，23个亚洲人和21个夏威夷/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护理点设备，该设备可以广泛传播给少数民族人口。

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