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Sökning: onr:"swepub:oai:DiVA.org:uu-326672" > Analytically Sensit...

LIBRIS Formathandbok  (Information om MARC21)
FältnamnIndikatorerMetadata
00004613naa a2200409 4500
001oai:DiVA.org:uu-326672
003SwePub
008170721s2017 | |||||||||||000 ||eng|
0247 a https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-3266722 URI
0247 a https://doi.org/10.1373/clinchem.2017.2718332 DOI
040 a (SwePub)uu
041 a engb eng
042 9 SwePub
072 7a ref2 swepub-contenttype
072 7a art2 swepub-publicationtype
1001 a Ebai, Tongeu Uppsala universitet,Science for Life Laboratory, SciLifeLab,Molekylära verktyg4 aut0 (Swepub:uu)toneb991
2451 0a Analytically Sensitive Protein Detection in Microtiter Plates by Proximity Ligation with Rolling Circle Amplification
264 c 2017-09-01
264 1b Oxford University Press (OUP),c 2017
338 a print2 rdacarrier
520 a BACKGROUND: Detecting proteins at low concentrations in plasma is crucial for early diagnosis. Current techniques in clinical routine, such as sandwich ELISA, provide sensitive protein detection because of a dependence on target recognition by pairs of antibodies, but detection of still lower protein concentrations is often called for. Proximity ligation assay with rolling circle amplification (PLARCA) is a modified proximity ligation assay (PLA) for analytically specific and sensitive protein detection via binding of target proteins by 3 antibodies, and signal amplification via rolling circle amplification (RCA) in microtiter wells, easily adapted to instrumentation in use in hospitals.METHODS: Proteins captured by immobilized antibodies were detected using a pair of oligonucleotide-conjugated antibodies. Upon target recognition, these PLA probes guided oligonucleotide ligation, followed by amplification via RCA of circular DNA strands that formed in the reaction. The RCA products were detected by horseradish peroxidase-labeled oligonucleotides to generate colorimetric reaction products with readout in an absorbance microplate reader.RESULTS: We compared detection of interleukin (IL)-4, IL-6, IL-8, p53, and growth differentiation factor-15 by PLARCA and conventional sandwich ELISA or immuno RCA. PLARCA detected lower concentrations of proteins and exhibited a broader dynamic range compared ELISA and iRCA using the same antibodies. IL-4 and IL-6 were detected in clinical samples at femtomolar concentrations, considerably lower than for ELISA.CONCLUSIONS: PLARCA offers detection of lower protein levels and increased dynamic ranges compared to ELISA. The PLARCA procedure may be adapted to routine instrumentation available in hospitals and research laboratories.
650 7a MEDICIN OCH HÄLSOVETENSKAPx Klinisk medicinx Klinisk laboratoriemedicin0 (SwePub)302232 hsv//swe
650 7a MEDICAL AND HEALTH SCIENCESx Clinical Medicinex Clinical Laboratory Medicine0 (SwePub)302232 hsv//eng
7001 a de Oliveira, Felipe Marques Souzau Uppsala universitet,Science for Life Laboratory, SciLifeLab,Molekylära verktyg4 aut0 (Swepub:uu)felma546
7001 a Löf, Lizau Uppsala universitet,Science for Life Laboratory, SciLifeLab,Molekylära verktyg4 aut0 (Swepub:uu)lizlo931
7001 a Wik, Lottau Uppsala universitet,Science for Life Laboratory, SciLifeLab,Molekylära verktyg4 aut0 (Swepub:uu)lowik226
7001 a Schweiger, Carolineu Charité Comprehensive Cancer Center, University of Berlin, Berlin, Germany; Institute of Pathology, Medical University of Graz, Graz, Austria;4 aut
7001 a Larsson, Andersu Uppsala universitet,Biokemisk struktur och funktion4 aut0 (Swepub:uu)andlarss
7001 a Keilholtz, Ulrichu Institute of Pathology, Medical University of Graz, Graz, Austria;4 aut
7001 a Haybaeck, Johannesu Charité Comprehensive Cancer Center, University of Berlin, Berlin, Germany; Department of Pathology, Otto von Guericke University Magdeburg, Magdeburg, Germany.4 aut
7001 a Landegren, Ulfu Uppsala universitet,Science for Life Laboratory, SciLifeLab,Molekylära verktyg4 aut0 (Swepub:uu)ulfland
7001 a Kamali-Moghaddam, Masoodu Uppsala universitet,Science for Life Laboratory, SciLifeLab,Molekylära verktyg4 aut0 (Swepub:uu)masookam
7102 a Uppsala universitetb Science for Life Laboratory, SciLifeLab4 org
7730 t Clinical Chemistryd : Oxford University Press (OUP)g 63:9, s. 1497-1505q 63:9<1497-1505x 0009-9147x 1530-8561
8564 u https://academic.oup.com/clinchem/article-pdf/63/9/1497/32647318/clinchem1497.pdf
8564 8u https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-326672
8564 8u https://doi.org/10.1373/clinchem.2017.271833

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