Low cost portable system for the rapid detection and drug resistance profiling of Tuberculosis in low resource environments

Tuberculosis TB is one of the major public health challenges in the world today with an estimated million deaths in More alarmingly the incidence of multi drug resistant MDR TB is rapidly increasing especially in low resource environments The goal of this project is to develop an inexpensive portable system and method using which it will be possible to both detect the presence of low loads of Mycobacterium tuberculosis Mtb in sputum samples AND perform Drug Susceptibility Testing DST for drug formulations all within days of sample collection The proposed approach involves a collecting all the Mtb cells present in ml of sputum sample into a small ul of buffer growth medium during the standard decontamination process using a set of super paramagnetic nanoparticles MNPs functionalized on the surface with amino polysaccharide moieties that bind to bacterial cells b dispensing the ul suspension into micro wells each containing different formulations of freeze dried growth media and or drugs of interest c monitor cells in each well using micro channel Electrical Impedance Spectroscopy m EIS m EIS reveals in real time if cells are growing dying or rendered static in the presence of antibiotic The first steps pre concentration using MNPs and loading into microfluidic channel will take less than hour and the monitoring of death growth stasis will take days In Phase I we will demonstrate proof of concept for the above approach by verifying the efficacy of our sample preparation pre concentration technique involving MNPs establishing the ability of the m EIS method to record in real time the death proliferation or statis of mycobacteria in the presence of candidate drug formulations and designing a prototype cassette containing wells and fabricating the same using D printing stereolithography In addition we will propose a detailed design electrical mechanical and fluidic of the automated system disposables and non disposable hardware that we will build in Phase II Being able to detect the presence of low loads of mycobacteria and gaining knowledge about the drug resistance profile of a particular isolate in days will be a game changer in our efforts to combat the spread of TB esp MDR TB It will lead to better outcomes for patients less mortality quicker recovery and also better public health less transmission to others and less fostering of drug resistance in populations Public Health Relevance Statement The World Health Organization has highlighted the emergence of Multi Drug Resistant MDR TB as a major global health challenge and stressed the need to expand rapid testing and detection of drug resistant TB cases The proposed instrument will cut the time needed to perform susceptibility tests against multiple drugs to days vs andgt weeks needed using current technology