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Sepsis – absolute quantification of MiRSeps-7 by using two-tailed RT-qPCR
University of Skövde, School of Bioscience.
2024 (English)Independent thesis Basic level (degree of Bachelor), 20 credits / 30 HE creditsStudent thesis
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. 

Place, publisher, year, edition, pages
2024. , p. 32
National Category
Biomedical Laboratory Science/Technology
Identifiers
URN: urn:nbn:se:his:diva-24462OAI: oai:DiVA.org:his-24462DiVA, id: diva2:1892892
Subject / course
Bioscience
Educational program
Bioscience - Molecular Biodesign
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Available from: 2024-08-28 Created: 2024-08-28 Last updated: 2024-08-28Bibliographically approved

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CiteExportLink to record
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