Dried blood sampling and digital readout to advance molecular diagnostics

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Fältnamn	Indikatorer	Metadata
000		05189nam a2200541 4500
001		oai:DiVA.org:uu-396325
003		SwePub
008		191103s2019 \| \|\|\|\|\|\|\|\|\|\|\|000 \|\|eng\|
020		a 9789151308104q print
024	7	a https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-3963252 URI
040		a (SwePub)uu
041		a engb eng
042		9 SwePub
072	7	a vet2 swepub-contenttype
072	7	a dok2 swepub-publicationtype
100	1	a Björkesten, Johan,d 1988-u Uppsala universitet,Science for Life Laboratory, SciLifeLab,Molekylära verktyg4 aut0 (Swepub:uu)johbj141
245	1 0	a Dried blood sampling and digital readout to advance molecular diagnostics
264	1	a Uppsala :b Acta Universitatis Upsaliensis,c 2019
300		a 60 s.
338		a electronic2 rdacarrier
490	0	a Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine,x 1651-6206 ;v 1615
520		a 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.
650	7	a MEDICIN OCH HÄLSOVETENSKAPx Medicinsk bioteknologi0 (SwePub)3042 hsv//swe
650	7	a MEDICAL AND HEALTH SCIENCESx Medical Biotechnology0 (SwePub)3042 hsv//eng
650	7	a NATURVETENSKAPx Biologix Biokemi och molekylärbiologi0 (SwePub)106022 hsv//swe
650	7	a NATURAL SCIENCESx Biological Sciencesx Biochemistry and Molecular Biology0 (SwePub)106022 hsv//eng
650	7	a TEKNIK OCH TEKNOLOGIERx Medicinteknikx Medicinsk laboratorie- och mätteknik0 (SwePub)206012 hsv//swe
650	7	a ENGINEERING AND TECHNOLOGYx Medical Engineeringx Medical Laboratory and Measurements Technologies0 (SwePub)206012 hsv//eng
653		a molecular diagnostics
653		a dried blood sample
653		a DBS
653		a digital readout
653		a digital enumeration
653		a DNA detection methods
653		a proximity extension assay
653		a protein stability
653		a genotyping
653		a rare mutations
653		a cell free DNA
653		a multiplex protein measurement.
653		a Farmaceutisk vetenskap
653		a Pharmaceutical Science
700	1	a Landegren, Ulf,c Professoru Uppsala universitet,Molekylära verktyg,Science for Life Laboratory, SciLifeLab4 ths0 (Swepub:uu)ulfland
700	1	a Högberg, Björn,c Professoru Karolinska institutet, department of medical biochemistry and biophysics4 opn
710	2	a Uppsala universitetb Science for Life Laboratory, SciLifeLab4 org
856	4	u https://uu.diva-portal.org/smash/get/diva2:1367388/FULLTEXT01.pdfx primaryx Raw objecty fulltext
856	4	u https://uu.diva-portal.org/smash/get/diva2:1367388/PREVIEW01.jpgx Previewy preview image
856	4 8	u https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-396325

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Av författaren/redakt...: Björkesten, Joha ...; Landegren, Ulf, ...; Högberg, Björn, ...

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MEDICIN OCH HÄLSOVETENSKAP: MEDICIN OCH HÄLS ...; och Medicinsk biotek ...

NATURVETENSKAP: NATURVETENSKAP; och Biologi; och Biokemi och mole ...

TEKNIK OCH TEKNOLOGIER: TEKNIK OCH TEKNO ...; och Medicinteknik; och Medicinsk labora ...

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