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- Björkesten, Johan,1988-Uppsala universitet,Science for Life Laboratory, SciLifeLab,Molekylära verktyg (author)
Dried blood sampling and digital readout to advance molecular diagnostics
- BookEnglish2019
Publisher, publication year, extent ...
- Uppsala :Acta Universitatis Upsaliensis,2019
- 60 s.
- electronicrdacarrier
Numbers
- LIBRIS-ID:oai:DiVA.org:uu-396325
- ISBN:9789151308104
- https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-396325URI
Supplementary language notes
- Language:English
- Summary in:English
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- SwePubSwePub
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Series
- Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine,1651-6206 ;1615
Notes
- A drastically increased capacity to measure large sets of molecular features in numerous patient samples in great detail will be required to fulfill the vision of precision medicine and wellness, which may characterize molecular diagnostics in the 21st century. Also sampling procedures need a renaissance to permit continuous sampling at population levels at reasonable cost.Blood sampling is typically performed via venipuncture to draw several milliliters of blood for plasma isolation. This is inconvenient, time-consuming and costly, as well as hard to standardize. The effect on plasma protein profiles by pre-centrifugation delay was investigated in Paper II, demonstrating time- and temperature-dependent release of proteins from blood cells upon delayed plasma isolation, but almost no protein degradation as analyzed by two 92-plex protein panels (Olink® Proteomics). An alternative sampling method, where blood drops from a finger stick are collected dried on paper, is relatively non-invasive, potentially home-based and cheap. Dried blood spots can also be shipped via regular mail and compactly stored. The effect of drying and long term storage stability of a large set of proteins from dried blood spots was investigated in Paper I using Olink® technology. The main findings were that drying slightly but consistently influenced the recorded levels of blood proteins, and that long-term storage decreased the detected levels of some of the proteins with half-lives of decades.Some molecular diagnostic investigations require great accuracy to be useful, arguing for digital enumeration of individual molecules. Digital PCR is the gold standard but Paper III presents an alternative approach based on rolling circle amplification of single molecules. Another instance where extreme assay performance is required is for rare mutation detection from liquid biopsies. Paper V presents a new method offering essentially error-free genotyping of individual molecules by majority-vote decisions for counting rare mutant DNA in blood. Yet other diagnostic investigations require very simple assays. Paper IV presents a novel one-step method to detect nucleic acid sequences by combining the power of rolling circle amplification and the specificity of DNA strand displacement in a format simple enough to be used at the point of care. Altogether, the thesis spans technologies for advanced molecular diagnostics, from sample collection over assay techniques to an improved readout.
Subject headings and genre
- MEDICIN OCH HÄLSOVETENSKAP Medicinsk bioteknologi hsv//swe
- MEDICAL AND HEALTH SCIENCES Medical Biotechnology hsv//eng
- NATURVETENSKAP Biologi Biokemi och molekylärbiologi hsv//swe
- NATURAL SCIENCES Biological Sciences Biochemistry and Molecular Biology hsv//eng
- TEKNIK OCH TEKNOLOGIER Medicinteknik Medicinsk laboratorie- och mätteknik hsv//swe
- ENGINEERING AND TECHNOLOGY Medical Engineering Medical Laboratory and Measurements Technologies hsv//eng
- molecular diagnostics
- dried blood sample
- DBS
- digital readout
- digital enumeration
- DNA detection methods
- proximity extension assay
- protein stability
- genotyping
- rare mutations
- cell free DNA
- multiplex protein measurement.
- Farmaceutisk vetenskap
- Pharmaceutical Science
Added entries (persons, corporate bodies, meetings, titles ...)
- Landegren, Ulf,ProfessorUppsala universitet,Molekylära verktyg,Science for Life Laboratory, SciLifeLab(Swepub:uu)ulfland (thesis advisor)
- Högberg, Björn,ProfessorKarolinska institutet, department of medical biochemistry and biophysics (opponent)
- Uppsala universitetScience for Life Laboratory, SciLifeLab (creator_code:org_t)
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