Utökning av panelför multiplex RT-MLPA av singel celler för prostatacancer diagnostik

Sammanfattning: The most common cancer form among men i prostate cancer and measurement of circulating tumor cells (CTCs) in the blood is a useful tool to evaluating the responset o treatment and progress of disease. CTC’s are cells that have detached from the original tumor and have spread in the blood sstem. In this master thesis a panel of 16 genes at single cell levels are analyzed using Reverse Transciptase Multiplexed Ligation-dependent Probe Amplification (RT-MLPA). The method uses gene specific reverse transcription primers to generate and amplify cDNA which MLPA probes are hybridized to. Correctly hybridized probes are ligated and amplified so that relative expression profiles can be calculated. One additional MLPA probe was designed to add to the existing panel. The results show that the MLPA reaction generates products from all genes in the panel when performed on synthetic MLPA prbe targets with equal concentrations. Results from totatl RNA on cell lines show that the reverse transcription and amplification of cDNA need further optimizations. When the whole assay is working it will be possible to evaluate gene expression from CTC’s that can help us understand the  progression and spread of prostate cancer in the body.