Högskolan i Skövde

his.sePublications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • apa-cv
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Future diagnostic of Sepsis: Biomarker miRSeps-5 manual vs robotic extraction and optimization of miRNA detection by Two-Tailed RT-qPCR technology
University of Skövde, School of Bioscience.
2022 (English)Independent thesis Basic level (degree of Bachelor), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

Sepsis is a syndrome developed in response to infection when a body’s immune system cannot adequately respond to infection. To this day, sepsis holds a peak position in mortality rates worldwide. Currently, SOFA score and blood culturing as means to diagnose sepsis are standard. However, blood culturing is lengthy and sepsis can develop at a faster rate. Faster diagnostic of sepsis in form of biomarker identification directly from the blood is one possible solution. One of the possible biomarker candidate is miRNA, a small non-coding RNA that participates in the regulation of many biological mechanisms of the human body. The aim of this work was to improve extraction of small RNA including miRNA from patient blood plasma via manual extraction using miRNeasy Advanced Kit (Qiagen) and robotic extraction using the same kit and QIAcube machine while comparing two methodologies. All samples containing extracted small RNA including miRNA were analysed with a Qubit 4.0 (Thermo Fisher Scientific) and DS-11+spectrophotometer (DeNovix). Furthermore, two-tailed RT-qPCR was investigated for potential to quantify synthetic and naturally occurring miRSeps-5 extracted from blood plasma. PCR System Machine was used to obtain Cq-values and melting curves. In order to use absolute quantification to quantify microRNA standard curve was optimized achieving a 116% efficiency and correlation coefficient R2=0.97. This work shows encouraging results of using two-tailed RT-qPCR to detect miRSeps-5 extracted from human plasma for future biomarker research. 

Place, publisher, year, edition, pages
2022. , p. 29
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:his:diva-21657OAI: oai:DiVA.org:his-21657DiVA, id: diva2:1684858
Subject / course
Bioscience
Educational program
Bioscience - Molecular Biodesign
Supervisors
Examiners
Available from: 2022-07-28 Created: 2022-07-28 Last updated: 2022-07-28Bibliographically approved

Open Access in DiVA

fulltext(1910 kB)307 downloads
File information
File name FULLTEXT01.pdfFile size 1910 kBChecksum SHA-512
850251c6a9e0c02087bb1b8f2cf2557c790c8ad709fb62aa55e932cf47c537167d9ffb669dc413073dc0ef52a0effb66be46376ab10e0b66f1640d5eca683677
Type fulltextMimetype application/pdf

By organisation
School of Bioscience
Biological Sciences

Search outside of DiVA

GoogleGoogle Scholar
Total: 318 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

urn-nbn

Altmetric score

urn-nbn
Total: 328 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • apa-cv
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf