Diagnosis of acute infections through microarray-based analysis of antibodies in lymphocyte supernatants

Reference Number: 
PGD5/CSande/Lymphocyte supernatants
Location: 
Kilifi
Country: 
Kenya
Supervisor: 
Dr Charles Sande, Dr Nelson Kibinge & Timothy Chege
Description: 

 

Background
Laboratory diagnosis of acute infections is often guided by clinical hypothesis formulated on the basis of the observed clinical symptoms. Since many infections result in largely overlapping symptoms, this approach is inherently imprecise and can result in long delays in establishing an accurate diagnosis. The majority of modern diagnostic assays are optimised for the detection of only a single antigen and often require several iterative trials to arrive at a definitive diagnosis. To improve diagnostic sensitivity, new efficient and multi-dimensional approaches are required. Serology is a well-established approach of identifying clinical infections. This method relies on the fact that most pathogens trigger a highly-specific antibody response, which can be measured as a proxy for the infection. However, classical serology is of limited clinical utility since antibodies take a few weeks to accumulate to detectable levels in blood. To address this problem we’ve recently reported on a new ELISA-based method for quantifying pathogen-specific antibodies produced by circulating immune-cells which can be detected within a few days of infection. In order to make this assay suitable for clinical diagnostics, we propose to increase its dimensionality and throughput by reformulating it into a protein-based microarray platform.

 

Aims of the project

  • To culture PBMCs from children admitted to hospital with acute infections and to isolate and concentrate antibodies from circulating plasmablasts
  • To detect and quantify antibodies to specific to different infectious pathogens using a protein microarray, spotted with ~80 proteins derived from highly prevalent infectious diseases
  • To validate the results of the microarraybased diagnosis using pathogen specific PCR

 

Skills to be acquired by the intern

  • Cell culture
  • Processing of microarraybased immunoassays
  • Primer design, conventional PCR, gel electrophoresis

 

Prefered training background of required intern

  1. BSc in any biological science
  2. Previous lab experience would be an advantage

 

Application deadline: 11 February 2019