CRISPR-TB for pediatric TB diagnosis and treatment response

PROJECT SUMMARY/ABSTRACT Children under 5-years of age account for ~50% of the 1.2 million new cases of pediatric TB each year, but are least likely to be diagnosed, and are at highest risk of death without prompt treatment. Young children are more likely to present with disseminated or extrapulmonary TB and paucibacillary disease, often missed by respiratory

sampling and our currently available diagnostics. Non-sputum diagnostic tools for TB detection and treatment response in young children, using easily obtained specimens are urgently needed. Our team has successfully developed an ultra-sensitive CRISPR-based approach (CRISPR-TB) that detects M. tuberculosis (Mtb) cell free DNA (cfDNA). In pilot evaluation with repository blood samples, CRISPR-TB

demonstrated high sensitivity (94%) and specificity (95%) among adults and children with TB and their asymptomatic household contacts in Eswatini. Among hospitalized Kenyan children with HIV (median age 2-years), CRISPR-TB detected 100% with microbiologically confirmed TB, and an additional 85% with clinically

diagnosed TB (i.e. missed by respiratory-based diagnostics). We now propose to expand CRISPR-TB evaluation in a large prospective cohort of 400 children with suspected TB (majority