Ändra sökning
RefereraExporteraLänk till posten
Permanent länk

Direktlänk
Referera
Referensformat
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Annat format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annat språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf
An 8 minute colorimetric paper-based reverse phase vertical flow serum microarray for screening of hyper IgE syndrome
KTH, Skolan för bioteknologi (BIO), Proteomik och nanobioteknologi.ORCID-id: 0000-0003-0344-8049
KTH, Skolan för bioteknologi (BIO), Proteomik och nanobioteknologi.
KTH, Skolan för bioteknologi (BIO), Proteomik och nanobioteknologi.
2015 (Engelska)Ingår i: The Analyst, ISSN 0003-2654, E-ISSN 1364-5528, Vol. 140, nr 21, s. 7327-7334Artikel i tidskrift (Refereegranskat) Published
Resurstyp
Text
Abstract [en]

Reverse phase microarrays are useful tools for affinity-based detection in hundreds of samples simultaneously. However, current methods typically require long assay times and fluorescent detection. Here we describe a paper-based Vertical Flow Microarray (VFM) assay as a rapid 8-minute colorimetric alternative for reverse phase microarray analysis. The VFM platform was optimized for detection of IgE with a detection limit of 1.9 μg mL-1 in whole serum. Optimized conditions were then used to screen 113 serum samples simultaneously for hyper IgE syndrome (hIgE), a rare primary immunodeficiency characterized by elevated levels of IgE. The same set of samples were then analysed with a conventional planar microarray with fluorescent detection for head-to-head testing. Both assays found elevated levels in three out of four hIgE patient samples, whereas no control samples displayed elevated levels in either method. The comparison experiments showed a good correlation between the two assays, as determined from a linear correlation study (Pearson's r = 0.76). Further, the assay-time reduction and reproducibility (intra assay CV = 12.4 ± 4.11%) demonstrate the applicability of the VFM platform for high throughput reverse phase screening.

Ort, förlag, år, upplaga, sidor
Royal Society of Chemistry, 2015. Vol. 140, nr 21, s. 7327-7334
Nationell ämneskategori
Kemi
Identifikatorer
URN: urn:nbn:se:kth:diva-181205DOI: 10.1039/c5an01013fISI: 000362795100031PubMedID: 26365343Scopus ID: 2-s2.0-84944096876OAI: oai:DiVA.org:kth-181205DiVA, id: diva2:902850
Forskningsfinansiär
Vetenskapsrådet
Anmärkning

QC 20160212

Tillgänglig från: 2016-02-12 Skapad: 2016-01-29 Senast uppdaterad: 2020-03-09Bibliografiskt granskad
Ingår i avhandling
1. Development of array systems for molecular diagnostic assays
Öppna denna publikation i ny flik eller fönster >>Development of array systems for molecular diagnostic assays
2018 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Abstract [en]

For molecular diagnostics, assays detecting biomarkers can be used to provide information to medical questions. Array formats such as microarrays and suspension bead arrays allow for multiplex assays, analyzing hundreds or thousands of analytes simultaneously in one single sample. There is a growing demand of multiplex assays, using panels of biomarkers, for the use in Point-of-Care (POC) diagnostic tests. These diagnostic tests require little or no equipment and relies on easy read-out systems. Lateral flow assays (LFAs) are well-known, paper-based POC assays, with advantageous features, which include performing the assay by capillary forces, stable reagents storage and naked-eye read-out. Vertical Flow Microarrays (VFMs) have previously been presented as an alternative to LFAs, circumventing downstream effects by vertically applying sample and reagents. In Paper I and II of this thesis, VFM arrays have been applied in the development of assays for molecular diagnosis, while Paper III explores Suspension Bead Array (SBA) format for biomarker discovery.

 

In Paper I, we have developed a DNA-VFM assay towards POC testing of Neisseria meningitidis, one of the major meningitis causing bacteria. Here, the target gene of N. meningitidis was amplified using Recombinase Polymerase Amplification (RPA). The amplified DNA was digested into ssDNA and hybridized to multiple VFM probes, for multiplex detection of different segments of the target gene. Optimization of the assay resulted in a Limit Of Detection (LOD) of 4.4 nM for amplification of synthetic DNA.

 

In Paper II, a VFM for reverse phase detection of IgE was developed. The assay was optimized using IgE spiked into IgE-negative serum, resulting in a LOD of 1.9 μg/mL. Optimized conditions were then used to screen a cohort of serum samples, including patients with rare primary immunodeficiency Hyper IgE syndrome and healthy controls. A comparative, traditional reverse phase assay was also performed on the same serum samples, showing comparable results to the reverse phase VFM.

 

In Paper III, a pediatric cohort from Plasmodium falciparum endemic Rwanda was analyzed using SBAs for proteins involved during various stages of malaria infection. Large, significant differences between cases and controls were found for 22 of the analyzed proteins. The majority of the candidate proteins presented validated previous work, nevertheless, several proteins were identified with no previously known link to malaria pathogenesis. Proteins discriminating between mild and severe malaria infection were also identified, showing minor separation between the two sample groups.

 

In Paper IV, a focus group discussion of using POC tests with Ugandan health care personnel was conducted. Health care personnel at different levels were interviewed about their perception of using POC tests in the health care system, including strengths and weaknesses. This study was designed to bridge some of the knowledge gap between the POC test developers and end-users.

Ort, förlag, år, upplaga, sidor
Stockholm: KTH Royal Institute of Technology, 2018. s. 71
Serie
TRITA-CBH-FOU ; 2018:5
Nationell ämneskategori
Medicinsk bioteknologi
Forskningsämne
Bioteknologi
Identifikatorer
urn:nbn:se:kth:diva-227159 (URN)978-91-7729-717-8 (ISBN)
Disputation
2018-05-25, Gardaulan, Folkhälsomyndigheten, Nobels väg 18, Solna, 10:00 (Engelska)
Opponent
Forskningsfinansiär
EU, Europeiska forskningsrådet, 615458
Anmärkning

QC 20180503

Tillgänglig från: 2018-05-03 Skapad: 2018-05-03 Senast uppdaterad: 2018-05-03Bibliografiskt granskad

Open Access i DiVA

Fulltext saknas i DiVA

Övriga länkar

Förlagets fulltextPubMedScopus

Personposter BETA

Reuterswärd, Philippa

Sök vidare i DiVA

Av författaren/redaktören
Reuterswärd, PhilippaGantelius, JesperAndersson Svahn, Helene
Av organisationen
Proteomik och nanobioteknologi
I samma tidskrift
The Analyst
Kemi

Sök vidare utanför DiVA

GoogleGoogle Scholar

doi
pubmed
urn-nbn

Altmetricpoäng

doi
pubmed
urn-nbn
Totalt: 149 träffar
RefereraExporteraLänk till posten
Permanent länk

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