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Sepsis – absolute quantification of MiRSeps-7 by using two-tailed RT-qPCR
Högskolan i Skövde, Institutionen för biovetenskap.
2024 (Engelska)Självständigt arbete på grundnivå (kandidatexamen), 20 poäng / 30 hpStudentuppsats (Examensarbete)
Abstract [en]

Sepsis is a life-threatening condition, which in time damages the patient’s organs. The quick sequence of events that could let a patient’s condition go from stable to severe in a few hours requires physicians to establish an early and accurate diagnosis. The research on new potential biomarkers is in full throttle. miRNA has elevated its potential as a diagnostic biomarker and may be included in a multiple-biomarker panel for sepsis diagnosis. In this project, the aim was first to evaluate if miRSeps-7 could be detected and quantified in human plasma by using the Two-tailed RT-qPCR method. Secondly, to compare the efficiency of two total RNA extraction methods (manual and robotic (Qiacube) extraction). This comparison was based on measuring the Hands-On Time (HOT) and Turnaround Time (TAT) during the extraction processes, along with analyzing the Cq values obtained from the qPCR reactions. Additionally, a QC check was performed on all samples after RNA extraction. No significant differences were discovered between the two extraction methods in terms of time management (e.g. HOT & TAT) nor the quality assessment, with quantification cycles (Cq). The small sample size was a major disadvantage. Amplification was achieved on 7/20 samples with the Two-tailed RT-qPCR. However, it has yet to be confirmed that the amplified product was miRSeps-7. 

Ort, förlag, år, upplaga, sidor
2024. , s. 32
Nationell ämneskategori
Biomedicinsk laboratorievetenskap/teknologi
Identifikatorer
URN: urn:nbn:se:his:diva-24462OAI: oai:DiVA.org:his-24462DiVA, id: diva2:1892892
Ämne / kurs
Biovetenskap
Utbildningsprogram
Biovetenskap - Molekylär biodesign
Handledare
Examinatorer
Tillgänglig från: 2024-08-28 Skapad: 2024-08-28 Senast uppdaterad: 2025-09-29Bibliografiskt granskad

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