November 14, 2019
By Benishar Kombut
The main purpose for this trip was to undergo 6 weeks training on the set up and performance of high through-put qPCR and ultra-sensitive qPCR for the diagnosis of low-density malaria infections, as well as to further develop my skills and expertise in conducting P. falciparum and P. vivax serological testing by ELISA and MagPix, data analysis and report/manuscript preparation. This was to be conducted at both the Burnet Institute and the Walter and Eliza Hall Institute.
During the 6 weeks, I was able to do the following activities/tasks:
- Bioplex assay were conducted at WEHI with Rhea Longley and Herbert Opi
– Watched and performed Bioplex assay on Magpix instrument using Thai clinical samples that Rhea and Herbert were working on. We tested for both total IgG and complement fixing(C1q) antibodies to 30 malaria proteins using this technique.
– Was able to set up a new protocol for this assay on the magpix instrument
– Transferred data output from magpix instrument into excel file and prism for data checking to ensure it was ok and ready for analysis
– Fortunate to watch the troubleshooting of magpix instrument when it became problematic during my training sessions
- Complement fixation assay with Liriye Kurtovic and Linda Reiling
– Tested for complement fixing(C1q) antibodies to MSP2 antigen using the ELISA technique
– Had a session on the importance of complement assays and why testing for complement is the appropriate functional antibody assay at the moment
– Had a session on tips for optimization of complement assays and general ELISA’s
- LAMP and RDT activities with Leanna Surrao
– Had the opportunity to see the various LAMP and RDT instrumentation developed by Axin that is being set up and optimized at the Burnet Institute
- Micronutrient ELISA assay with Eliza Davidson
Performed ELISA assay on serum from women in the Healthy Mothers, Healthy Babies cohort to optimize two micro nutrient assay kits;
– Retinol Binding protein ELISA kit
– Folic Acid (Vitamin A) ELISA kit
- qPCR and quality control testing of high sensitive RDT kits for malaria with Shazia Ruybal
– Was able to watch and perform qPCR for Plasmodium vivax species on malaria in pregnancy samples according to protocol
– Performed quality control testing of high sensitive malaria rapid diagnostic test kits using culture panels provided by FIND
- Manuscript writing
– Was able to discuss some of my previously generated data with Joanne Chan and Shazia Ruybal. They provided constructive feedback on how to analyse my data and materials on how to be able to compile a manuscript.
– Produced an outline of data analysis approach and conducted analysis accordingly
– Attended an Introductory course to R statistical analysis software
- Other activities
– I had the opportunity to present our recent completed work on the high sensitivity RDT for malaria in pregnancy in PNG to the Burnet Board Members and also at the Burnet Student Symposium. I was also privileged to take part in an interview by GSK Australia, for Professor Brendan Crabb’s GSK award in malaria research.
The ACREME travel award has given me the opportunity to develop expertise in molecular high through-put qPCR and magpix for serological testing. It has provided me both the experience and exposure of working in an international environment and in world class laboratories, using cutting edge tools. It also enabled me to establish relationships and network with both Burnet and WEHI staff in Melbourne. I am so grateful to have undergone this training through this award and would like to sincerely acknowledge ACREME for giving me this invaluable opportunity.