Detection of IL28B SNP DNA from Buccal Epithelial Cells, Small Amounts of Serum, and Dried Blood Spots

Background & Aims: Point mutations in the coding region of the interleukin 28 gene (rs12979860) have recently been identified for predicting the outcome of treatment of hepatitis C virus infection. This polymorphism detection was based on whole blood DNA extraction. Alternatively, DNA for genetic diagnosis has been derived from buccal epithelial cells (BEC), dried blood spots (DBS), and genomic DNA from serum. The aim of the study was to investigate the reliability and accuracy of alternative routes of testing for single nucleotide polymorphism allele rs12979860CC. Methods: Blood, plasma, and sera samples from 200 patients were extracted (400 mL). Buccal smears were tested using an FTA card. To simulate postal delay, we tested the influence of storage at ambient temperature on the different sources of DNA at five time points (baseline, 48 h, 6 days, 9 days, and 12 days) Results: There was 100 % concordance between blood, plasma, sera, and BEC, validating the use of DNA extracted from BEC collected on cytology brushes for genetic testing. Genetic variations in HPTR1 gene were detected using smear technique in blood smear (3620 copies) as well as in buccal smears (5870 copies). These results are similar to those for whole blood diluted at 1/10. A minimum of 0.04 mL, 4 mL, and 40 mL was necessary to obtain exploitable results respectively for whole blood, sera, and plasma. No significant variation between each time point was observed for the different sources of DNA. IL28B SNPs analysis at these different time points showed the same results using the four sources of DNA

0243: Changes in mitral regurgitation severity after successful aortic valve implantation: impact on the development of heart failure

Backgroundthe association of severe aortic stenosis and mitral regurgitation (MR) is not rare in patients undergoing transcatheter aortic valve implantation (TAVI). The effect of TAVI on MR and its impact on outcome was evaluated.MethodsAll patients undergoing TAVI in our center were evaluated. Patients with paravalvular leak or residual stenosis after TAVI were excluded. Ninety out of 166 patients undergoing TAVI had follow-up data on post-procedural MR and constituted the patient population. Mitral regurgitation was graded on a 0-4 scale and divided into two categories for survival analysis, minimal (none/mild MR [grade 0-1/4]) and significant MR (moderate /severe MR [grade 2-4/4]). Patients were followed for a median of 427 days for death and development of heart failure (HF).ResultsAverage age was 81±6; 49% were male and the average Euroscore was 22±15. Mitral regurgitation before TAVI was minimal in 52 pts (58%) and significant in 38 pts (42%). Post TAVI, MR was minimal in 56 pts (62%) and significant in 34 pts (38%). Sixteen pts (18%)developed worsening MR with an increase of at least one MR grade post TAVI, 25 pts (28%) has an improvement in the MR (MR grade was less) and 49 pts (54%) no change occurred in the MR. Development of significant MR post successful TAVI was a strong predictor of the development of HF (49±9% vs 70±6%, respectively, p=0.02) as well as the combined end point of death and HF (event free survival 46%±9% versus 64%±7%).ConclusionsMR post TAVI without aortic regurgitation or residual stenosis is a commmon issue and predicts the developemnt of HF and death