High-throughput qRT-PCR validation of blood microRNAs in XXXX .

@entity1 @entity1149 @entity2 @entity1030
UNASSIGNED: Validation of biomarkers is essential to advance the translational process to clinical application. Although there exists an increasing number of reports on small non-coding RNAs (microRNAs) as minimally-invasive markers from blood, serum or plasma, just a limited number is verified in follow-up studies. We used qRT-PCR to evaluate a known miRNA signature measured from blood that allowed for separation between @entity1 with @entity1149 ( @entity1149 ), @entity1030 and healthy controls.From the data of our previous microarray studies we selected a panel of 235 miRNAs related to @entity2 and @entity1030 . We observed a high concordance between the AUC values of our initial microarray screening and the qRT-PCR data (correlation of 0.704, p < 10-16). Overall, 90.3% of markers were successfully validated. Among the top markers that were concordant between both studies we found hsa-miR-20b-5p, hsa-miR-20a-5p, hsa-miR-17-5p, and hsa-miR-106a-5p. The qRT-PCR analysis also confirmed that @entity1149 @entity1 could be accurately differentiated from unaffected controls: a subset of five markers was sufficient to separate @entity1149 @entity1 from unaffected controls with accuracy of 94.5% (specificity and sensitivity of 98% and 91%). Beyond differentiation from controls, we also succeeded in separating @entity1149 @entity1 from @entity1 with @entity1030 . MiRNAs that were identified as relevant for the separation between @entity2 and @entity1030 by both qRT-PCR and the array-based studies included hsa-miR-26a-5p, hsa-miR-328-3p and hsa-miR-1224-3p. Although for differentiation between @entity1149 @entity1 from @entity1030 @entity1 more markers were required, still high accuracy rates were obtained (5 markers: 78.8%; 10 markers: 83.9%; 50 markers: 87.6%).
