Evaluation of Manual and QIAcube miRNA Extraction from Plasma with the miRNeasy Advanced Kit from Qiagen

Sammanfattning: Sepsis is a syndrome causing millions of deaths every year. The diagnosis today uses several parameters one of which is blood culture. It can take up to 48 hours to get results. Sepsis can lead to major organ dysfunction in a span of four hours; therefore time is of the essence. Diagnosis of sepsis using multiple biomarkers a “multi marker panel” is currently being studied. One parameter suggested to be included is miRNA. There is need for an efficient extraction method to get the miRNA from the patients’ blood. This can be done by using a kit either manually or in a machine such as the QIAcube. The miRNeasy Serum/Plasma Advanced Kit (Qiagen) was used in both a manual and a QIAcube miRNA extraction with two volumes of plasma (200 μl and 100 μl) from self-assessed healthy individuals. There were 10 extractions made with each method and volume, in total 40 extractions was performed. The miRNA concentration was measure with a Qubit and purity was measured with a nanodrop. The miRNA concentration was statistically significantly higher using the QIAcube 100 μl compared to the other volume and the manual extractions. The turn-around time was shorter with the manual extraction while the hands-on time was shorter in the QIAcube methods. In conclusionthere is still research to be done on both miRNA and miRNA extraction before it can be included in a multi marker panel for sepsis. However the QIAcube does show potential to be used in a clinical laboratory.