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Tuberculosis
Tuberculosis (TB) is an infectious disease that is caused by the Mycobacterium tuberculosis (M.tb) species of bacteria (1). This disease dates back to Neolithic times where it was previously referred to as Pott spondylitis, scrofula and consumption (2). While evidence shows that the disease has been in existence for over 10000 years (3-5), the causative agent of TB was only discovered by Robert Koch in 1882 (1).
Despite this being discovered already for over 100 years the disease continues to cause epidemics worldwide. It was estimated that approximately 11 million people in Africa were killed by the disease between the year 2000 and 2016. While the mortality rate (regarding TB) remains high in Africa, one should take note that the disease is a worldwide problem. The estimated TB incidence rates can be seen in Figure 1 (6).
Figure 1 illustrates the incidence rate of the people infected with TB worldwide in 2016. Just as TB is a problem, there are a large number of people in the world that are infected with the human immunodeficiency virus (HIV). This disease has the ability to alter an individual’s immune system and often these patients then contract TB also. In the year 2016 the World Health Organisation (WHO) reported that approximately 1.1 million people were living with a TB and HIV co-infection worldwide (6).
This total contributes largely to the incidence rate of TB around the world. While the TB-HIV co-infection is a major problem worldwide it was reported in 2016 that more people were killed by TB disease than HIV alone (6). TB is a growing epidemic and is expected to expand if the disease is not curbed soon.
Chapter 1: Introduction
1.1.Tuberculosis
1.1.1. Disease progression in the host
1.1.2. Active TB versus Latent TB
1.2. Treatment of Tuberculosis
1.3. Diagnosis of TB
1.4. Blood-based diagnosis
1.5. Mycolic acids
1.6. Immunochromatographic based diagnostics
1.7. Problem Statement
1.8. Hypothesis
1.9. Aims
Chapter 2: Aims and strategies of the study
Chapter 3: Development of antibody displacement probe
3.1. Introduction
3.2. Materials and Methods
3.2.1. Screening of library for MA binders
3.2.2. Isolation of scFvs
3.2.3. Evaluation of binding as scFvs
3.2.4. Plasmid isolation and purification
3.2.5. Plasmid amplification
3.2.6. Agarose gel electrophoresis
3.2.7. Restriction enzyme digestion
3.2.8. DNA Precipitation
3.2.9. Crystal violet gel
3.2.10. Ligation
3.2.11. Transformations
3.2.12. Colony PCR
3.2.13. Sequencing
3.2.14. Resuscitation of HEK cell
3.2.15. Transfections
3.2.16. Zeocin selection
3.2.17. Gallibody functionality
Chapter 4: Manuscript
Chapter 5: Concluding Discussion
