Research Projects

Researcher: Dr JG Nel

The contribution of pneumolysin and hydrogen peroxide to the induction of neutrophil extracellular traps by Streptococcus pneumoniae 
 

Neutrophils are vital for defence against microbial pathogens. They are generated in great number in the bone marrow and transiently circulate in blood. Neutrophils respond to infection by phagocytosing and killing microbial pathogens. The killing of pathogens is accomplished through the use of an arsenal of toxic molecules represented by proteolytic enzymes, reactive oxygen species (ROS), and an array of bactericidal proteins.

 

Researcher: Prof R. Pool, Dr JJC Potgieter and Mrs Andrea Prinsloo

A randomized, cross over, open label trial to evaluate the pharmacokinetics, efficacy and safety profile of a recombinant Factor VIII product in previously treated subjects with severe haemophilia

A recombinant Factor VIII product will be evaluated using potency determinations (chromogenic assay per European pharmacopeia versus chromogenic assay label adjusted to factor VIII:c in plasma one stage coagulation assay potency) as measured by bleeding rate. The trial will aslo evaluate the safety, tolerability and efficacy of the product. The potential for inhibitory antibody formation during prophylactic treatment with this agent will also be assessed.  
 

Researcher: Prof R. Pool, Dr JJC Potgieter and Mrs Andrea Prinsloo

A phase II/III, randomized, cross-over, open label trial to demonstrate superiority of prophylaxis over on-demand therapy in previously treated subjects with severe haemophilia A treated with plasma protein free recombinant FVIII formulated with sucrose

The primary objective of this trial is to demonstrate the superiority of prophylaxis over on-demand therapy by a clinically significant decrease in bleeding rate following 12 months of treatment with the study drug. Secondary objectives are to demonstrate the superiority of prophylaxis versus on-demand therapy, to evaluate the safety of the study drug and to assess its potential to induce the formation of inhibitory antibodies.

 

Researcher: Dr SF van Rooyen

The effect of temperature on the stability of Protein C and Protein S assays in thawed plasma

This study will report on the protein C and protein S assay stability for thawed samples stored at different temperatures for 24 hours.   Samples will be collected from normal volunteers, as well as volunteers with known low values of protein C and S. These samples will be prepared within two hours from collection, according to the laboratory’s standard operating procedure for obtaining plasma from citrated whole blood.

 

Researcher: Dr W Gouws

Comparing one stage- and chromogenic factor VIII assays in the diagnosis of patients with haemophilia A

The one stage method of determining the factor VIII level in haemophilia A patients is currently the method of choice at Steve Biko Academic Hospital`s core laboratory. As proven in the literature the factor VIII level measured in mild haemophilia A patients is lower with the chromogenic assay than the one stage assay. The severity of the haemophilia A is underestimated when using the one stage assay and this directly influences the management of these patients.

In this prospective study the factor VIII levels determined with the one stage clotting assay will be compared to the factor VIII level measured with the chromogenic assay.

 

Researcher: Dr E Beltchev

Evaluation of the AMLprofiler® (acute myeloid leukaemia profiler) compared to the standard diagnostic procedures in patients with acute myeloid leukaemia.

The aims of the proposed study will be to evaluate the benefits of the AMLprofiler® in the South African context. The project will compare the current diagnostic procedures performed routinely in patients with acute myeloid leukaemia to that of the AMLprofiler®. The variables to be compared will include cost, concordance between results obtained using current techniques and AMLprofiler® and time required to execute the tests.

 

Researcher: Dr G George

A method for establishing the presence of bcr-abl in patients with chronic myeloid leukaemia using flow cytometry

Flow cytometry allows for the identification of particles based on attributes such as size and fluorescence intensity. A Cytometric Bead Array (BM™) system provides a way of coupling a soluble analyte with beads of known size or fluorescence thus making it possible to detect analytes by flow cytometry. This technique will be employed to detect bcr-abl usion proteins in human blood research samples.

 

Researcher: Mrs A Prinsloo

Specific bcr-abl mutations in chronic myeloid leukaemia patients

Micro-RNAs (miRNAs) have recently been identified as a class of small functional non-coding RNAs of 18-25 nucleotides (nt) which bind to the 3’ untranslated region (UTR) of mRNAs down-regulating gene expression at a post-transcriptional level. These RNA fragments play critical roles in cellular processes including metabolism, apoptosis, differentiation and development. Micro-RNAs are expressed in specific haematological cell types with a regulatory function in early haematopoietic differentiation, erythropoiesis, granulopoiesis, megakaryopoiesis as well as lymphoid development. This is a pilot study to determine if the miR-17-92 polycistron is expressed in vivo with the objectives of using quantitative reverse transcriptase (qRT)-PCR to determine expression of miRNA in CML patients and healthy individuals.

 

Researcher: Dr T Chetty

Epidemiological investigation of HIV-related lymphoma in the South African population

The proposed project plans to obtain epidemiological data on HIV-related lymphoma in South Africa. More specifically, this will be a retrospective evaluation of both the incidence and prevalence of HIV-related lymphomas in the Gauteng province (initially) over the past 3 years (2009 to 2011). The rationale for this component of the study stems from the fact that our group is involved in a project that aims to develop an HIV resistant immune system by means of generating CCR5 null haematopoietic stem cells for bone marrow transplantation with the objective of curing HIV/AIDS. Along with collaborators in Geneva and Zurich, Switzerland, the project has progressed beyond proof of concept in vitro and is currently entering evaluation in a humanised mouse model. Once the pre-clinical phase has been completed, clinical trials will be initiated in South Africa based on the high prevalence of patients with HIV-related lymphoma. These patients have been selected since many of them require bone marrow transplantation following chemotherapy. However, there is to date no reference epidemiological data on this population in South Africa. It is therefore critical that this population be accurately defined so as to ensure that the most appropriate sub-population be selected for the planned clinical trial.

 

Researcher: Ms M Mokgathi

Method validation of an automated cell counter for CSF analysis

The Advia 2120® cell counter has an FDA approved automated CSF assay as part of its routine test repertoire. A series of CSF specimens will be analysed in terms of the red cell count, white cell count and white cell differential. Factors such as linearity, carry over and within run precision and accuracy will be assessed.

 

Researcher: Ms D Matlebjane:

Utility of the reticulocyte haemoglobin content in the diagnosis of iron deficiency in pregnant women

Determination of the reticulocyte hemoglobin content (CHr) provides an early measure of functional iron deficiency because reticulocytes are the earliest red cell precursorsreleased into blood and circulate for only 48 hours before maturing into erythrocytes. The clinical utility of CHr for the diagnosis of the anemia of chronic disease has not been carefully studied. This project will look at the utility of the CHr, together with other traditional haematological and biochemical parameters in establishing the diagnosis of iron deficiency in pregnancy.

 

Researcher: Ms S Aboobaker

Paroxysmal nocturnal haemoglobinuria diagnosis by flow cytometry using the FLAER method

PNH is an acquired hematopoietic stem cell disorder leading to a partial or absolute deficiency of all glycophosphatidyl-inositol (GPI)-linked proteins. FLAER is an inactive variant of aerolysin that that retains specificity for GPI-linked structures but does not cause lysis of cells. The goal of this study is to develop a FLAER-based assay to diagnose and monitor patients with PNH and to improve detection of minor populations of PNH clones.

 

Researcher: Ms A Kemp

The establishing a reference range of leucocytes in CSF of healthy individuals using the Advia 2120® automated cell counter 

Normal CSF may contain up to 5 WBCs per mm in adults and 20 WBCs per mm in newborns. Cerebrospinal fluid will be obtained from patients undergoing spinal anaesthesia and the specimens will be analysed using the Advia 2120® haematology analyser to establish normal cell values for CSF at our institution

 

Researcher: Ms T Kalua

Using pre-warmed RPMI and an increased number of washes to improve kappa and lambda visualisation through flow cytometry

The purpose of this study is to ascertain whether either pre-warmed RPMI medium or increased number of washes positively influences the visualisation of the kappa and lambda light chains on the flow cytometer.

 

Researcher: Ms R Jansen  

Evaluation of real time PCR to detect Prothrombin and Factor V Leiden mutations

The aim of this study is to evaluate a real time PCR assay for possible use in molecular diagnostics to detect both the Prothrombin G20210A and Factor V Leiden mutations