Högskolan i Skövde

his.sePublikasjoner
Endre søk
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • apa-cv
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf
Comparison of manual and semi-automatic RNA extraction methods using two-tailed RT-qPCR for absolute quantification as part of the sepsis diagnosis research
Högskolan i Skövde, Institutionen för biovetenskap.
2023 (engelsk)Independent thesis Basic level (degree of Bachelor), 20 poäng / 30 hpOppgave
Abstract [en]

Nowadays, sepsis has become a major healthcare problem. Its variance of symptoms and the lack of time to act makes it greatly difficult to treat. An early diagnosis using biomarkers, particularly miRNA, could potentially increase the patient’s prognosis as well as reduce the use of antibiotics for the treatment. The lack of method optimization for miRNA extraction and quantification calls for investigation prior to the construction of a multi-biomarker panel for sepsis diagnosis. The aim of this project was to examine and compare manual and semi-automatic extraction methodologies through the small RNA quantity and RNA quality, as well as test the detection and quantification abilities of the novel technique, two-tailed RT-qPCR. 30 extractions have been performed, their extracted elutions have been subjected to quality and quantity control and detection and absolute quantification through the two-tailed RT-qPCR. The results show no significant differences between the quantity and quality of the RNA extracted using both methods. Time management, on the contrary, reported significant differences between the two methods. On the other hand, the two-tailed RT-qPCR successfully amplified the miRNA candidate from as little as 100 µL of healthy plasma. The absolute quantification showed the miRNA candidate’s low concentration in plasma. Moreover, the qPCR efficiency was irregular during the project which may alert of contamination or unspecific primers. However, the melt curve showed a single amplicon which suggests great specificity. The detection and quantification of the miRNA candidate have been successful, though further investigation is recommended.

sted, utgiver, år, opplag, sider
2023. , s. 37
HSV kategori
Identifikatorer
URN: urn:nbn:se:his:diva-22891OAI: oai:DiVA.org:his-22891DiVA, id: diva2:1777840
Fag / kurs
Bioscience
Veileder
Examiner
Merknad

Utbytesstudent

Tilgjengelig fra: 2023-06-30 Laget: 2023-06-30 Sist oppdatert: 2023-06-30bibliografisk kontrollert

Open Access i DiVA

fulltext(1484 kB)205 nedlastinger
Filinformasjon
Fil FULLTEXT01.pdfFilstørrelse 1484 kBChecksum SHA-512
3274df3b72c8fc4330de91bca5b795797c1b422e5ab595d278925bda0ec1dad65764e6e3a2f1e2c41ef72959a72f94944948fe822c766b6c776f8744ebbfcfe7
Type fulltextMimetype application/pdf

Av organisasjonen

Søk utenfor DiVA

GoogleGoogle Scholar
Totalt: 205 nedlastinger
Antall nedlastinger er summen av alle nedlastinger av alle fulltekster. Det kan for eksempel være tidligere versjoner som er ikke lenger tilgjengelige

urn-nbn

Altmetric

urn-nbn
Totalt: 436 treff
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • apa-cv
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf