 Welcome everyone. I am Adzab Erzma from the Erasmus M.C. University Medical Center in Rotterdam and I want to tell you about our recent publication in human mutation entitled Identification and Characterization of Abherent GAA Pre-MRNA splicing in Pompa disease using a generic approach. In our lab we are working on research involving the process of splicing with a focus on Pompa disease. Like in Pompa disease many monogenetic disorders can be caused by genetic variants which change native splicing. However in diagnostics these variants are often not functionally characterized and sometimes even missed. We hypothesized there should be a fast and easy method to essay aberrant splicing events using existing PCR techniques. Here we show that such an essay is feasible, is able to detect aberrant splicing event and that it is time and cost effective. The method is implemented as follows. The essay consists of two parts. In the first part here you look at qualitative differences in splicing which can be detected by flanking action PCR. The products are separated on a gel and when an aberrant splicing occurs there will be an extra band which can be analyzed by sequencing. In the second part we can quantify this aberrant splicing by QPCR using primers within each axon. By applying this method you can detect whether there is a splicing variant and then you can identify that. You can also quantify the extent of aberrant splicing and you can correlate that with disease severity. We applied this method to patients with pulmonary disease who had a partial or uncharacterized variant. Here you see a couple of those patients and here are the variants. For example we detect several aberrant splicing events but also leaky wild-type splicing. Here we have a full intron 10 inclusion. We can also have a partial inclusion of intron 10 etc. The wisest method interesting. First it is important for genetic counseling so family members can be tested for the same variant. Second the patient often wants to confirm the diagnosis on the DNA level. Third it is important for future patients to know whether the splicing variant is pathogenic or not and finally one could predict disease severity by looking at the extent of aberrant splicing. We invite you to test this approach on your gene of interest and we hope you enjoy reading the article.