The purpose of this study was to investigate whether the bacterial RNA detected by polymerase chain reaction (PCR) and reverse transcription (RT)-PCR methods in middle ear effusion (MEE) for pediatric chronic otitis media with effusion (OME) originated from live bacteria. Degradation of RNA was observed by spectroscopic analysis; we also investigated the effect of MEE on the digestive activity of RNase. The optical density of RNA solution was stable within 3 h. MEE could not degrade the RNA, while RNase could rapidly digest the RNA.
Blood samples are used as a biological source to discover biomarkers of hematological and non-hematological disorders. The present study shows the impact of different experimental conditions associated with cell lysis buffer, TRI-reagent protocol and blood cell storage buffer and their correlation with the quantity, quality and Adrenomedullin gene expression levels of total RNA when RT-PCR technique is used. A leukocyte cell bank protocol is also proposed for further mRNA expression analysis using RNAlater as storage buffer.
