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EMBO

Workshop on molecular diagnostics of tick-borne diseases (EMBO/ICTTD)

The aim of the microarray part of the workshop was to use a custom oligonucleotide microarray platform to determine what parasites were present in unknown samples. These samples could have contained any number of parasites, which makes microarrays a useful detection tool to use since it has great capacity to screen many potential hits in parallel.

Unique DNA samples (16-22mer oligonucleotides), representing each of the parasites that were screened for, were spotted onto glass slides. These slides were then hybridised to a labelled mixture made from the same samples that were tested with the reverse-line blot technique (RLB). Fluorescent detection of the hybridised spots revealed the presence of specific parasites in the unknown sample that was being screened. The microarray results were then compared to those obtained with RLB. These two methods were quite consistent but the microarray platform could still be further optimised, mainly to increase sensitivity while decreasing non-specific hybridisation. The two images below each show the result of a tested sample and indicate the presence of different parasites. They can be identified according to their position on the array, with four replicates per sample.
 

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