Konference: 2015 XI. Dny diagnostické, prediktivní a experimentální onkologie
Téma: Biomarkery nádorových onemocnění II
Číslo abstraktu: 038
Autoři: Mgr. Hanuš Slavík; MUDr. Josef Srovnal, Ph.D.; Mgr. Zuzana Bačová, Ph.D.; Ing. Rastislav Slavkovský, Ph.D.
Introduction
Exhaled breath condensate (EBC) is a rich and easily accessible source of biomarkers of many lung diseases. Its main advantage is the possibility of non-invasive collection. EBC mainly contains volatile organic and inorganic molecules but also proteins and nucleic acids. MiRNA could be a candidate tumor and nontumor biomarker of lung diseases because of its stability and presence in EBC.
Materials/methods
22 samples from healthy individuals have been collected yet (11 men and 11 women aged 23 to 37 years old). RNA from these samples was isolated using Urine microRNA Purification Kit (Norgen). Concentrations were measured with an Agilent Bioanalyzer 2100. Various options of miRNA profiling were tested: RT-qPCR - MiRNA qPCR Panels (Exiqon), microarrays - MiRNA 3.0 Array (Affymetrix) and NGS - True Seq Small RNA (Illumina).
Results and conclusions
Only small amount of miRNA was possible to extract from EBC samples (average total yield was 3.91 ± 1.63 ng from 1-2 ml EBC). Simple RT-qPCR showed the presence of abundant miRNAs. RT-qPCR arrays seem to be the best option for EBC analysis because only 3 ng of small RNAs are sufficient as an input. Microarrays are less useful because fluorescent signals from individual probes are very weak and the recommended input is about 100 ng. Despite of this facts, used samples passed the quality control. NGS seems to be inappropriate for these analyses, because it was not possible to prepare cDNA libraries from such a small amounts of RNA.
IGA LF UP 2015_10 and TAČR TE02000058
Datum přednesení příspěvku: 3. 12. 2015
