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A novel assay for the detection of Hepatitis C virus in blood plasma, using padlock probes and rolling circle amplification
University of Skövde, School of Bioscience.
2023 (English)Independent thesis Basic level (degree of Bachelor), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

Hepatitis C viral infection is a globally widespread blood-borne disease affecting the liver, causing cirrhosis and hepatocellular carcinoma. This type of liver cancer is mainly caused by chronic Hepatitis C and Hepatitis B. Specific detection with the following correct treatment is crucial to reduce the overall burden of the disease. This work focused on investigating whether the padlock probes and rolling circle amplification can detect Hepatitis C, determining the limit of detection, and if any blood components would inhibit the reactions. All oligonucleotides were tested for ligation functionality in 10% TBE-Ureal gel electrophoresis and used with rolling circle amplification and phi29 polymerase to determine if eye read-out was possible. The lowest concentration of detection was found to be 10 pM. To avoid inhibition in blood plasma, samples were pre-treated at 95 ˚C for five minutes. Eye read-out was possible after amplification, with 30% plasma at the highest and 5% plasma at the lowest in samples. In conclusion, this novel assay using padlock probes, a detection oligonucleotide, and rolling circle amplification holds promise in developing a simplified new detection technique for the diagnostics of Hepatitis C.

Place, publisher, year, edition, pages
2023. , p. 40
National Category
Medical Bioscience
Identifiers
URN: urn:nbn:se:his:diva-22927OAI: oai:DiVA.org:his-22927DiVA, id: diva2:1778442
External cooperation
Uppsala Universitet
Subject / course
Bioscience
Educational program
Bioscience - Molecular Biodesign
Supervisors
Examiners
Available from: 2023-07-02 Created: 2023-07-02 Last updated: 2023-07-02Bibliographically approved

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CiteExportLink to record
Permanent link

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