TB Diagnostics at the Point of Care

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.