As part of the CATTEDRA Interreg project, Experteam has the role of developing a new in vitro diagnostic test (IVD) for the characterization of immune-mediated pathologies in pediatric patients. This is a complex process that requires an in-depth study of the many genes involved in the JAK-STAT signaling pathway. Currently, Experteam has developed a new mRNA extraction and sequencing protocol to identify genes associated with certain inflammatory diseases.
The extraction phase takes place using the Nuetraction Nucleic Acid Purification System BFEX-32 automatic extractor, and is followed by the quantification of the amount of RNA obtained and the analysis of its integrity. The latter is evaluated using the RIN (RNA Integrity Number) algorithm, which assigns a value from 1 (completely degraded) to 10 (intact) to each sample and generates a characteristic graph of the total RNA population (Figure 1).
The transcriptome amplification and sequencing phase uses a Next-Generation Sequencing technology known as 3' RNA-Seq, which allows for the analysis of only the coding RNA sequences and quickly obtain a complete expression profile for each sample.
Through the study of the gene expression profile of different patients performed by Experteam, it will be possible to process the fundamental data to identify some of the genes associated with complex inflammatory diseases and, finally, to develop an effective and innovative diagnostic system.
Image: Graphs of the integrity of extracted mRNA. The graphs are indicative of a fair level of integrity, the major peaks corresponding to 28S and I8S ribosomal RNAs.
Operation co-funded by the European Regional Development Fund
