Team:IISER Pune/Projects
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Project Background
Tuberculosis, an infectious pulmonary disease, is caused by Mycobacterium tuberculosis which spreads through air. The most common symptoms include cough lasting more than 3 weeks, weakness, fever, chest pain, coughing up blood/sputum, etc. The minimum duration of treatment is 6 months. If not treated properly, this disease can be fatal. According to the WHO Global TB Report 2014, India has the highest TB incidence. A large population combined with poor hygienic conditions, moderate temperatures, socio-economic conditions- all contribute to the high prevalence of TB in India.
The average duration from the appearance of symptoms to the beginning of treatment is 2-4 months. The reason for this large gap is that M. tuberculosis is a slow growing pathogenic bacterium with a doubling time of approximately 24 hours in tissues. This property of M. tuberculosis hinders rapid detection in sputum samples obtained from patients. Also, the commonly used diagnostic techniques in India not only require a time period of 2-4 weeks but also cannot detect lower concentrations of mycobacteria in the sample from the patients.
Thus, there is a need for a diagnostic technique that is cost effective, yet is quick and can still detect mycobacteria in lower concentrations.
Project Overview
Tuberculosis is caused by Mycobacterium tuberculosis. In India, TB poses a great health threat with nearly 40% Indians having either latent or active TB. The most commonly used TB diagnostic techniques in India require a minimum of 2-3 weeks, thus causing a delay in treatment. Some of the techniques used require high-end instruments which many clinical labs may not have access to. Mycobacterium tuberculosis is a slow-growing bacteria and hence, the detection of this bacteria in patient samples using present techniques requires a longer duration. In the quest to improve TB diagnostics, we propose a technique that does away with the need of high-end instruments and complicated procedures. This technique aims to give an easy-to-identify output upon the addition of our genetic device to the sample.
Our genetic device consists of 3 modules - Hijack, Detection and Termination.
Hijack: This module of our project aims at hijacking the cell cycle to increase the growth rate of cells.The hijacking is achieved by means of a genetic oscillator that targets 2 proteins - dnaA and ftsZ.
Detection: As the number of cells increase, this module aims at detecting these cells by producing a colour output. For this purpose, three possibilities were explored - chromoproteins, enzyme-substrate reactions and carotenoid production in cells.
Terminator: Once the Hijack machinery takes over, the cells divide rapidly as their growth rate increases. The Terminator turns on the suicide switch at a particular cell density and prevents uncontrolled proliferation of these cells.