Detection of exosomal mirna from different volumes of biofluids as biomarkers for the diagnosis of sepsis : Future diagnostics of sepsis

Abstract: Sepsis, a life-threatening condition which results from a dysregulation of host response to infection and leads to multiple organ dysfunction, is a cause for great concern. The current gold standard of detection – Blood culturing – is a highly time-consuming process and so, research has proposed the use of biomarkers. Current biomarkers, C-reactive protein and Procalcitonin, though good indicators, individually show certain limitations with respect to the specificity and sensitivity. Hence, as a step forward from singleplex biomarkers, the development of a multi-marker panel was suggested. For this purpose, the use of microRNAs (miRNAs) were employed to serve as potential biomarkers for the detection of sepsis. The aim of this study was to determine whether a higher concentration of miRNA would be obtained from a larger volume of plasma as well as to see if the miRNA present in blood can be used for the diagnosis of sepsis. Extractions were carried out using the QIAGEN exoRNeasy Plasma: Midi & Maxi Kits from plasma and Norgen’s Total RNA Purification Kit from blood. The samples were analysed and quantified using the Qubit® microRNA assay kit & Qubit® 3.0 Fluorometer and the NanoDrop™ 2000 Spectrophotometer. Statistical analysis of the results revealed that there was a significant difference between miRNA concentrations in the two volumes of plasma analysed. Based on the accurate Qubit measurements and readings, it was concluded that a larger volume of plasma, does yield a higher concentration of miRNA. In addition, it was also established that the miRNA detected in blood, could be used as probable biomarkers for the diagnosis of sepsis.