Magister Curationis - MCur / Multidrug resistant TB (MDR-TB) is a form of TB caused by bacteria (germs) that are resistant to the usual drugs that are used to treat "normal" TB. The duration of treatment for MDR-TB is a maximum of 22 months. People with MDR-TB are treated in specialized tertiary hospitals and in out-patient clinics in the PHC facilities. The treatment includes a six months injectable phase with a wide range of TB drugs. The adverse effects of MDR-TB drugs are among the worst side effects ever reported by patients. The aim of the current study was to explore the experiences of adverse effects of MDR-TB treatment amongst patients in a primary health care facility in the Western Cape. An explorative qualitative study design was used to explore the experiences of patient with the adverse effects of MDR-TB treatment in a primary health care facility in the Western Cape. In depth interviews were conducted with 12 MDR-TB patients. Data analysis was done by using the Tesch's method of content analysis. The study revealed that participating MDR-TB patients experienced various emotional, financial, physical and social challenges. Participants explained that the experience of being on MDR-TB treatment is emotionally draining; the pain and discomfort of the adverse effect of treatment makes a person to feel anxious and depressed. Financially they depended on social grants because they had to stop working after starting treatment. They could not function well physically because of the toxic nature of the adverse effects of treatment; which resulted in fatigue, dizziness and burning sensation on the feet and hands. They were faced with a lot of stigma from the community and even family members because of their illness. The study also revealed that in spite of the challenges and obstacles the participants were all motivated to complete their treatment and get cured. It is recommended that more support structures be made available for patients who are being treated for MDRT-TB such as; psychotherapy, social support and counselling on health education. Provision needs to be made for patients who are receiving daily injection; for it to be given in their homes. Health care providers treating MDR-TB patients need to do home visits together with MDR-TB adherence counsellors, to monitor the physical wellbeing of patients at home. This will also provide patients with the platform to discuss their health concerns in a more accommodative and relaxed environment. New drug regimen with fewer tablets and less treatment duration is needed for MDR-TB.

Mycobacterium tuberculosis has caused a resurgence in pulmonary disease in both developed and developing countries in recent times, particularly amongst people infected with the human immunodeficiency virus. The disease has assumed epidemic proportions in South Africa and in the Eastern Cape Province in particular. Of further concern is the isolation of increasing numbers of multiply drug resistant strains. Knowledge of the genetic capability of this organism is essential for the successful development of novel antibiotics and vaccines in an attempt to bring the global pandemic under control. Measurement of the genetic diversity of the organism may significantly contribute to such knowledge, and is of vital importance in monitoring epidemics and in improving treatment and control of the disease. This will entail answering a number of questions related to the degree of genetic diversity amongst strains, to the difference between urban and rural strains, and between drug resistant and drug sensitive strains, and to the geographical distribution of strains. In order to establish such baseline information, RAPD profiling of a large population of isolates from the western and central regions of the Eastern Cape Province was undertaken. A smaller number of drug resistant strains from a small area of KwaZulu-Natal were also analysed, with a view to establishing the genetic difference between strains from the two provinces. Cluster analysis, analysis of molecular variance and Geographical Information Systems technology were used to analyse the RAPD profiles generated. An unexpectedly high degree of genetic diversity was detected in strains from both provinces. While no correlation was seen between genetic diversity and either urban-rural situation or geographical location, a small degree of population structure could be correlated with drug resistance in the Eastern Cape. Furthermore, a significant degree of population structure was detected between strains from the two provinces, although this was still within the parameters for conspecific populations. Future work is necessary to further characterise strains from rural areas of both provinces, as well as from the eastern region of the Eastern Cape in an attempt to pinpoint the cause of the separation of the provincial populations.

Two of the primary occupational health threats to employees in the mining industry are noise-induced hearing loss (NIHL) and occupational lung diseases (OLD) with Tuberculosis (TB) included in the latter. The objective of this study was to investigate the hearing profile of a group of gold miners with and without TB to determine the effect of TB and its associated risk profile on hearing. Workers in AngloGold Ashanti mine in South Africa were recruited due to the fact that they present with these two health threats namely NIHL and TB. The audiological and medical surveillance data of 2698 subjects (between the years 2001 and 2009) were used in analyses. Hearing thresholds for the air conduction frequencies (0.5, 1, 2, 3, 4, 6, 8 KHz) in both ears were analysed in conjunction with biographic and occupational data. Subjects were divided into three groups, two experimental groups (Single TB treatment, n= 911 and Multiple TB treatment, n= 376) and one control group (n= 1411). A highly significant difference (p<0.01) was noted between the control group and both TB treatment groups across most frequencies and hearing parameters analysed, although the higher frequencies were more affected. Pair wise comparisons revealed the largest differences in hearing thresholds throughout between the control group and the multiple TB treatment groups. The smallest differences in hearing thresholds were evident between the two TB groups with the multiple TB treatment group presenting with the poorest thresholds. TB and its related risk profile had a pronounced influence on the decline of hearing thresholds. Thresholds for the multiple TB treatment group indicated more deterioration than the hearing thresholds of the single TB treatment group. This may point to the possibility that the influence of repeated TB on the subjects’ hearing thresholds over time was more pronounced than a single incidence of TB. It is still necessary however to separate the effects of the disease from the effects of the treatment on hearing. / Dissertation (MCommunication Pathology)--University of Pretoria, 2012. / Speech-Language Pathology and Audiology / Unrestricted

Thesis (M.Sc.(Microbiology)) -- University of Limpopo, 2022 / This study aimed to investigate antimycobacterial and cytotoxic compounds from Cassia petersiana. Cassia petersiana was selected for the current study based on its traditional use for treating tuberculosis (TB) symptoms. Extraction is an important step in the use of medicinal plants; hence, solvents of varying polarity were employed to extract a wide range of compounds where chloroform was the best extractant (67 mg). As there is no relation between the amount of plant material extracted and the bioactivity of the extracts, standard tests were used to determine the presence of different phytochemical constituents from Cassia petersiana and the total phenolic, flavonoid, and tannin contents were quantified using colorimetric assays. It was revealed that all the tested phytochemical constituents were present, and it was proven that phenolic compounds were the most abundant, followed by the tannins, while the flavonoids were the least among the common phytochemical constituents quantified. The phytochemical compounds were further profiled on thin-layer chromatography (TLC) and developed in BEA, CEF, and EMW solvent systems. Colourful compounds which indicated diverse phytochemicals were visualised with both vanillin-sulphuric acid and ultraviolet light on the phytochemical chromatograms and good separation of the compounds was from the BEA solvent system. The qualitative and quantitative antioxidant activity and antimycobacterial activity assays were used to evaluate the extracts from Cassia petersiana. Minimal antioxidant activity was observed on the qualitative antioxidant activity profile. These findings correlated with the minimal quantity of antioxidants from extracts of Cassia petersiana from the quantitative antioxidant assays; ferric reducing power and DPPH scavenging activity assays. Cassia petersiana extracts had bioactivity against Mycobacterium smegmatis as indicated by the lowest MIC value. The cell viability effects of the acetone crude extract from Cassia petersiana were evaluated against the tryptophan hydroxylase-1 (TPH-1) macrophage cells. Large scale extraction procedure was employed to extract a sufficient amount of plant material in preparation for the isolation of the bioactive compound. Bioassay-guided fractionation combined with column chromatography and TLC were used to isolate and purify the bioactive compound from the n-hexane extract of Cassia petersiana. The purified isolated compound was elucidated as β-sitosterol, which showed remarkable bioactivity against Mycobacterium smegmatis only on the TLC-bioautographic assay, while the quantitative antimycobacterial activity was higher xx with the MIC value of 2.5 mg/mL. Although β-sitosterol is known as a good antioxidant, it showed no antioxidant activity on the qualitative antioxidant activity assay. Therefore, further studies, including in vivo assay, are recommended on the isolated compound to evaluate its biological activities before consideration of its use in the development of alternative drugs.

The purpose of this study was to explore and describe the personal experiences of enrolled nurses while caring for patients infected with multidrug-resistant tuberculosis (MDR-TB) in an urban tuberculosis hospital in KwaZulu-Natal province, South Africa. Generic qualitative research was conducted with a sample of purposively selected enrolled nurses who cared for MDR-TB patients. Data was collected through in-depth individual interviews and analysed using Colaizzi’s (1978) method of data analysis. The research findings revealed six major themes: the working context, fear of contracting the disease, problems that have an impact on the quality of nursing care, nurses' perceptions of the patients, support structures and nurses' expressed needs. The findings of this study indicate that the nurses work in a challenging environment and need to be supported, as they experience more negative than positive feelings while caring for these patients. / Health Studies / (M.A. (Health Studies))

Submitted in fulfilment of the requirements for the Degree in Masters of Technology in Nursing, Durban University of Technology, 2012. / Background : The current alarming growth of diseases and complications, especially in Africa, makes the integration of traditional and conventional health practices a priority in medical training, research and planning, and the funding of health services. Unplanned and/or unintended treatment non-compliance and unnecessary deaths from diseases like tuberculosis and Human Immunodeficiency Virus are escalating in spite of health information and/or education, support groups and awareness events. The World Health Organisation recommends Directly Observed Treatment Strategy for illnesses like tuberculosis, and suggests the inclusion of traditional health practitioners in the strategy because they are constantly in contact with the community and could therefore be utilized as reminders, support system, doctors and care givers. Therefore it is a high priority that traditional health practitioners be integrated into partnership with conventional medicine practitioners, as they are considered the entry point to primary health care programmes in South Africa. Aim of the study The aim of this study was to explore the nature of the partnership between the African traditional and conventional health care in the eThekwini District. Methodolody : A qualitative, multiple case study design was used to explore the partnership between African traditional and conventional health care within the South African health care system in the eThekwini district of KwaZulu-Natal Province. In attempting to explore and understand the extent to which both these health care systems work together, a qualitative research method was used. All ethical issues were considered after which individual interviews were conducted using an interview guide and a tape recorder. A cross-case synthesis was used to analyse data. Results : Results from the study suggest that a partnership is far from being implemented by both the Traditional Health Practitioners and Conventional Health Care Practitioners. It is apparent that they both do not share a common vision. The government has some responsibility and a major role to play in guiding such a partnership and making sure that the South African community is provided with best practices governed by policies and legislation that are transparent, fair and legally binding to everybody involved.

The purpose of this study was to analyse the tracking systems used to identify patients with tuberculosis who missed a clinic appointment at primary health care facilities in Waterberg District. A quantitative descriptive correlation design was used to determine if there was an association between the tracking systems used and the defaulter rates. Data was collected using a questionnaire with nurses from 46 primary health care facilities, defaulter rates were accessed from the ETR.Net and the two sets of data were correlated. The results showed that the blue folder yielded the lowest mean defaulter rate while the green card yielded the highest mean defaulter rate. Nurses were unaware of the true defaulter rate in their facilities as they underestimated these rates. They therefore did not implement relevant intervention strategies to recall patients or find ways to improve the tracking systems used to reduce the defaulter rate in their health facilities. / Health Studies / M.A. (Health Studies)

Thesis (MScMedSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: Setting This study was conducted in the Tygerberg district, Cape Town, in the Western Cape, South Africa Background The evaluation of early tuberculosis (TB) treatment response is based on month 2 sputum culture status. This method of evaluation has a number of limitations: the test requires relatively advanced laboratory infrastructure and procedures, it takes several weeks to obtain results and is a relatively a poor marker at predicting treatment response. The discovery of potential host markers which reflect the efficacy of early treatment would be of great importance for clinical management of individual patients. The treatment failure would be detectable earlier than at week 8 of treatment. The duration of clinical trials of new anti-tuberculosis drugs may also be substantially reduced by such markers if these would be measurable earlier than at week 8 of therapy. Objectives 1) To evaluate diluted, 7-day whole blood cultures stimulated with live Mycobacterium tuberculosis (M.tb) for the presence of host markers of early TB treatment response 2) To evaluate an overnight, undiluted, M.tb antigen stimulated whole blood culture Quantiferon Gold In Tube (QFT-GIT) supernatants for host markers of early TB treatment response The study designs were as follows: In study one, baseline samples and samples from week 1, week 2 and week 4 of treatment from 30 cured TB patients were selected from a larger biomarker study, in which whole blood was stimulated with live M.tb or left unstimulated. Fifty seven host markers were measured in supernatants by multiplex cytokine arrays. In study two, baseline samples and samples from week 2 and week 8 of treatment from 19 cured TB patients were randomly selected from the placebo group in a micronutrient supplement study. QFT-GIT supernatants from these participants were assessed through multiplex cytokine arrays for levels of fifty seven host markers. All of the participants in both studies were Human Immunodeficiency Virus (HIV) negative. Changes in marker expression over time and between fast and slow responders to treatment were evaluated. Comparability between the two culture methods was assessed for markers that were evaluated in both studies. Results In study one, the majority of host markers showed significant changes over time in the unstimulated supernatants. Only GRO and IL-1beta changed significantly in an antigen-specific manner (background levels subtracted). No significant changes were observed between fast and slow responders. In study two, the majority of host markers showed significant changes over time in the unstimulated supernatants whereas only MDC and IL-4 changed during the observation period in antigen stimulated levels. Significant differences were observed between fast and slow responders at pre-treatment for IL-13 Ag-Nil and IL-1betaAg-Nil . Conclusion This study revealed, antigen-specific responses showed only limited potential for early TB treatment response monitoring, but may have potential in differentiating between treatment outcomes. Future investigations may have to include later time points during treatment as these were not included in the present assessment. The QFT-GIT samples do not appear to be equivalent to live M.tb stimulated 7-day whole blood assays. / AFRIKAANSE OPSOMMING: Instelling Die studie is uitgevoer in die Tygerbergdistrik, Kaapstad, Wes-Kaap, Suid-Afrika. Agtergrond Die evaluering van die respons op vroeë tuberkulose (TB) behandeling word gebaseer op die status van maand 2 sputum kulture. Hierdie evalueringsmetode het ‘n paar beperkinge: die toets benodig relatief gevorderde laboratorium infrastruktuur en prosedures, die toetsuitslae is eers na ‘n paar weke beskikbaar en dit is n relatiewe swak merker om repons op behandeling te voorspel. Die ontdekking van potensiële selfmerkers wat die doeltreffendheid van vroeë behandeling weerspieël sal van groot belang wees vir die kliniese bestuur van individuele pasiënte. Mislukking van die behandeling sal sodoende voor week 8 van behandeling waargeneem kan word. Die tydsduur van kliniese proewe van nuwe anti-tuberkulose medikasie mag ook baie verkort word met sulke merkers as dit voor week 8 van behandeling gemeet kan word. Doelwitte 1) Om verdunde, 7-dae oue volbloedkulture, met lewende Mikobakterium tuberkulosis (M.tb) gestimuleer, te evalueer vir die teenwoordigheid van vroeë TB behandeling respons selfmerkers. 2) Om die supernatant van oornag, onverdunde, M.tb antigeen gestimuleerde volbloedkulture Quantiferon Gold In Tube (QFT-GIT) vir vroeë behandeling respons selfmerkers te evalueer. Die studie-ontwerpe was soos volg: Met studie een is basislynmonsters en monsters verkry na week 1, week 2 en week 4 van behandeling van 30 geneesde TB-pasiënte geselekteer uit ‘n groter biomerkerstudie waarin die volbloed met lewende M.tb gestimuleer is of ongestimuleer gelaat is. Sewe-en-vyftig selfmerkers is in die supernatante gemeet deur middel van multipleks sitokine arrays. Met studie twee is basislynmonsters en monsters verkry na week 2 en week 8 van behandeling van 19 geneesde TB-pasiënte lukraak uit die plasebo-groep in ‘n mikrovoedingstowwe-aanvullingstudie geselekteer. Vlakke van 57 selfmerkers is in die QFT-GIT supernatante van hierdie deelnemers, deur middel van die multipleks sitokine arrays, bepaal. Al die deelnemers van beide studies was HIV negatief. Veranderinge in merker-uitdrukking oor tyd, asook tussen vinnige en stadige respons tot behandeling, is ge-evalueer. Die vergelykbaarheid van die twee kultuurmetodes is geassesseer ten opsigte van die ge-evalueerde merkers in albei studies. Resultate Met studie een het die meerderheid van die selfmerkers in die ongestimuleerde supernatante kenmerkende verandering oor tyd gewys. Slegs GRO en IL-1beta het aansienlik verander in die antigeenspesifieke wyse (agtergrond vlakke afgetrek). Geen kenmerkende veranderinge was waargeneem tussen die vinnige en stadige respons pasiënte nie. Met studie twee het die meerderheid van die selfmerkers aansienlike veranderinge oor tyd in die ongestimuleerde supernatante gewys, in vergelyking waar net die MDC en IL-4 veranderinge gedurende die observasie periode in antigeen gestimuleerde vlakke getoon het. Kenmerkende verskille is tussen die vinnige en stadige respons pasiënte in voorbehandeling vir IL-13 Ag-Nil en IL-1betaAg-Nil waargeneem. Gevolgtrekking Die studie bewys dat antigeenspesifieke response slegs beperkte potensiaal vir vroeë TB behandeling respons monitering het, maar mag die potensiaall vir onderskeidende behandeling uitkomste hê. Toekomstige ondersoeke sal dalk latere tydpunte gedurende die behandeling moet insluit aangesien dit nie in hierdie evaluasie ingesluit is nie. Die QFT-IT monsters verskyn nie as gelykwaardig met die lewendig M.tb gestimuleerde 7-dae volbloed toetse nie.

Thesis (PhD)--Stellenbosch University, 2012. / Includes bibliography / ENGLISH ABSTRACT: Mycobacterium tuberculosis, the causative organism of tuberculosis (TB), is a major cause for mortality and morbidity world-wide with a death toll only second to HIV among infectious diseases. Drug resistance is widespread and cases of multiple drug resistant TB (MDR-TB) and extensively drug resistant TB (XDR-TB) have emerged in several countries. Drug treatment is problematic and new drugs are not developed rapidly enough to offset the rapid drug resistance mutation rate of M. tuberculosis. Simple and effective diagnostics are required to contain the spread of the disease as current routine diagnostics are not fulfilling this role. Additionally, current rapid TB diagnostics are out of reach to resource poor settings due to infrastructure, cost and skill requirements. Novel TB diagnostics are thus required that meet these requirements. Mycobacteriophages are phages that infect mycobacteria and could offer a viable and cost effective alternative rapid TB diagnostics. In this study, an affinity-tagged fluorescent reporter mycobacteriophage is described, which was engineered to act as a TB diagnostic. Its performance proved favourable and superior to current existing mycobacteriophage-based TB diagnostics. / AFRIKAANSE OPSOMMING: Mycobacterium tuberculosis, die organisme verantwoordelik vir tuberkulose (TB), is `n groot bron van mortaliteit en morbiditeit wêreldwyd en slegs HIV is verantwoordelik vir groter getalle sterftes as gevolg van n aansteeklike siekte. Middelweerstandigheid is algemeen en gevalle van meervoudigemiddelweerstandige tuberkulose (MDR-TB) en uiters weerstandige tuberkulose (XDR-TB) kom in verskeie lande voor. Antibiotika behandeling is problematies en nuwe anti-TB middels word nie vinnig genoeg ontwikkel om die antibiotika weerstandigheid mutasie spoed van M. tuberculosis te bekamp nie. Doeltreffende diagnostiese toetse word benodig om die verspreiding van die siekte te beheer en bestaande roetine diagnostiese toetse voldoen tans nie aan hierdie vereiste nie. Behalwe hiervoor, is huidige vinnige TB diagnostiese toetse buite bereik van arm instansies weens vereistes aan infrastruktuur, meegaande kostes en werknemervaardigheid. Nuwe TB diagnostiese toetse is dus nodig om aan hierdie vereistes te voldoen. Mikobacteriofaage is fage wat mikobacteria infekteer en kan moontlik 'n lewensvatbare en koste-effektiewe alternatief bied vir vinnige TB diagnostiese toetse. In hierdie studie word 'n affiniteitgekoppelde fluoreserende rapporteringsmikobakteriofaag beskryf wat ontwerp is om op te tree as `n nuwe vinnige TB diagnostiese toets. Die werking hiervan vertoon gunstige en beter resultate as die huidige, mikobacteriofaaggebaseerde TB-diagnostiese toetse.

Thesis (MScMedSc)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: Anti-tuberculosis drugs target specific essential cellular processes and structural components. The first line drug, rifampicin (RIF) is a RNA polymerase inhibitor which targets the β-subunit and subsequently inhibits the initiation of transcription. Previous proteomic and transcriptomic analyses have shown that exposure to RIF for 24hrs significantly increased the abundance of proteins involved in energy metabolism in clinical isolates. No studies have been done to describe the transcriptional responses to RIF in an in vitro RIF resistant M. tuberculosis isolate. Application of in vitro mutants is novel since it will exclude most of the confounding factors which may be present in clinical isolates obtained from patients where the bacterium may have been incubated for several weeks or even years. This study aimed to determine the effect of prolonged exposure to RIF and the effect of the rpoB Ser531Leu mutation on the expression of energy metabolism genes, sigma factors and a regulator in RIF mono-resistant in vitro mutants with different levels of RIF resistance (minimum inhibitory concentration (MIC): 40μg/ml and 70μg/ml). RIF mono-resistant in vitro mutants were generated from a pan susceptible Beijing cluster 208 progenitor using the Luria Delbruck assay. In vitro RIF mono-resistant mutants harbouring the Ser531Leu rpoB mutation and which displayed different levels of RIF resistance were selected. To assess the effect of prolonged RIF exposure on the expression of candidate genes, the in vitro mutants were cultured in liquid media and exposed to RIF for 1, 7 and 14 days. High quality RNA was extracted from these cultures at each time point and Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR) was done on the selected candidate genes. The results indicate that limited expression of energy metabolism genes and sigma factors was observed after prolonged RIF exposure. In addition, the activity of the regulator (Rv1846c) was down-regulated in the presence of RIF explaining the up-regulated state of energy metabolism genes. To assess the effect of the rpoB Ser531Leu mutation on the candidate genes, RNA was extracted from the RIF unexposed culture at mid-log phase. RT-qPCR was done for each in vitro mutant in addition to the wild-type progenitor isolate. These results show that energy metabolism genes and sigma factors were significantly up-regulated in the RIF resistant mutantss harbouring an rpoB Ser531Leu mutation. This suggests that the mutation had a significant effect on the cellular energy cost due to the up-regulated state of the energy metabolism genes. In addition, an increase in the expression of sigma factors may be required to compensate for the rpoB mutation by enforcing the binding of the RNA polymerase and sigma factors to the promoter for transcription to be initiated. It is therefore important to assess these candidate genes for their potential as novel candidates for future drug design as this is an important aspect to influence tuberculosis control. / AFRIKAANSE OPSOMMING: Teen-tuberkulose middels teiken essensiële sellulêre prosesse en strukturele komponente. Die eerste linie teen-tuberkulose middel, rifampisien (RIF) is ʼn RNS polimerase inhibeerder wat die β-subeenheid teiken en daarna die inisiasie van transkripsie onderdruk. Vorige proteomiese en transkriptomiese analises het getoon dat blootstelling aan RIF vir 24 uur beduidende styging in sekere protiene wat verband hou met energie metabolisme in kliniese isolate veroorsaak. Die huidige studie poog om die effek van langdurige RIF blootstelling, asook die effek van die rpoB Ser531Leu mutasie op die uitdrukking van energie metabolisme gene, sigma faktore en reguleerders op RIF-enkel weerstandige in vitro mutante by verskillende vlakke van RIF weerstandigheid (Minimum Inhiberende Konsentrasie (MIK): 40μg/ml en 70μg/ml) te ondersoek. RIF-enkelweerstandige in vitro mutante isolate is gegenereer van ʼn sensitiewe Beijing 208 stamfamilielid deur die Luria Delbruck metode. In vitro RIF enkelweerstandige mutante met die rpoB Ser531Leu mutasie en verskillende vlakke van RIF weerstandigheid is geselekteer. Om die langdurige effek van RIF blootstelling op kandidaat geen uitdrukking te ondersoek, is in vitro mutante isolate gegroei in vloeibare medium en blootgestel aan RIF vir 1, 7 en 14 dae. Goeie kwaliteit RNS is geëkstraheer van hierdie kulture by elke tydpunt om Werklike-tyd Kwantitatiewe Polimerase Ketting Reaksie (RT-qPCR) op die kandidaat gene uit te voer. Die resultate toon dat ʼn beperkte aantal energie metabolisme en sigma faktor gene uitgedruk was na RIF blootstelling. Verder is die uitdrukking van die reguleerder (Rv1846c) af gereguleer in die teenwoordigheid van RIF en dit verduidelik die op gereguleerde energie metaboliese geen patroon. Om die effek van die rpoB Ser531Leu mutasie op die kandidaat gene te evalueer, is RNS geëkstraheer van ʼn weerstandige en RIF sensitiewe kultuur wat nie blootgestel was aan RIF nie. RT-qPCR is uit gevoer op elke in vitro mutante isolaat asook op ʼn sensitiewe isolaat sonder ʼn mutasie. Hierdie resultate toon dat energie metabolisme gene en sigma faktore beduidend opreguleer word in die isolate met ʼn rpoB Ser531Leu mutasie. Dit dui daarop dat die mutasie ʼn beduidende effek op die sellulêre energie koste het, omdat die energie metabolisme gene op gereguleer is. Verder kan ʼn toename in die uitdrukking van sigma faktore benodig word om die effek van die rpoB mutasie te oorkom deur binding van die RNS polimerase en die sigma faktore aan die promotor om transkripsie inisiasie te forseer. Dit is daarom belangrik om hierdie kandidaat gene verder te ondersoek vir toekomstige ontwikkeling van teenmiddels teen tuberkulose.