Sökning: onr:"swepub:oai:DiVA.org:kth-282251" >
Sub-attomole detect...
Sub-attomole detection of HIV-1 using padlock probes and rolling circle amplification combined with microfluidic affinity chromatography
-
- Soares, Ruben R. G. (författare)
- Stockholms universitet,KTH,Science for Life Laboratory, SciLifeLab,Nanobioteknologi,Stockholm Univ, Dept Biochem & Biophys, Sci Life Lab, Solna, Sweden.,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab),KTH Royal Institute of Technology, Sweden
-
- Varela, João C. (författare)
- Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab),Stockholm Univ, Dept Biochem & Biophys, Sci Life Lab, Solna, Sweden.
-
- Neogi, Ujjwal (författare)
- Karolinska Institutet
-
visa fler...
-
- Ciftci, Sibel (författare)
- Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab),Stockholm Univ, Dept Biochem & Biophys, Sci Life Lab, Solna, Sweden.
-
- Ashokkumar, Manickam (författare)
- Karolinska Inst, Dept Lab Med, Div Clin Microbiol, Huddinge, Sweden.
-
- Pinto, Ines Fernandes (författare)
- KTH,Nanobioteknologi,Science for Life Laboratory, SciLifeLab
-
- Nilsson, Mats (författare)
- Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab),Stockholm Univ, Dept Biochem & Biophys, Sci Life Lab, Solna, Sweden.
-
- Madaboosi, Narayanan (författare)
- Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab),Stockholm Univ, Dept Biochem & Biophys, Sci Life Lab, Solna, Sweden.
-
- Russom, Aman, Assoc.Prof. (författare)
- KTH,Nanobioteknologi,Science for Life Laboratory, SciLifeLab
-
visa färre...
-
(creator_code:org_t)
- Elsevier BV, 2020
- 2020
- Engelska.
-
Ingår i: Biosensors & bioelectronics. - : Elsevier BV. - 0956-5663 .- 1873-4235. ; 166
- Relaterad länk:
-
https://urn.kb.se/re...
-
visa fler...
-
https://doi.org/10.1...
-
https://urn.kb.se/re...
-
http://kipublication...
-
visa färre...
Abstract
Ämnesord
Stäng
- Despite significant progress in diagnostics and disease management during the past decades, human immunodeficiency virus (HIV) infections are still responsible for nearly 1 million deaths every year, mostly in resource-limited settings. Thus, novel, accurate and cost-effective tools for viral load monitoring become crucial to allow specific diagnostics and the effective monitoring of the associated antiviral therapies. Herein, we report an effective combination of a (1) padlock probe (PLP)-mediated rolling circle amplification (RCA) bioassay and an (2) agarose bead-based microfluidic device for the affinity chromatography-based capture and detection of RCA products (RCPs) pre-labelled simultaneously with biotin and an organic fluorophore. This method allowed the efficient capture of similar to 1 mu m-sized RCPs followed by their quantification either as discrete signals or an average fluorescence signal, thus being compatible with both high-resolution imaging for maximum sensitivity as well as simpler optical detection setups. A limit of detection < 30 fM was obtained for HIV-1 synthetic target with just a single round of RCA, comparable to recently reported procedures requiring technically complex amplification strategies such as hyperbranching and/or enzymatic digestion/amplification. Furthermore, targeting a set of five conserved regions in the HIV-1 gag gene, the method could specifically detect HIV-1 in 293T cell culture supernatants, as well as a set of 11 HIV-1 NIH reference samples with four different subtypes. The reported method provides simplicity of operation, unique versatility of signal transduction (i.e. average or discrete signals), and potential coupling with previously reported miniaturized photodetectors. These combined features hold promise for bringing RCA-based molecular diagnostics closer to the point-of-care.
Ämnesord
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
- NATURVETENSKAP -- Biologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences (hsv//eng)
Nyckelord
- Rolling circle amplification
- Streptavidin-biotin
- Beads
- HIV
- Microfluidics
- Concentration
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
Hitta via bibliotek
Till lärosätets databas
- Av författaren/redakt...
-
Soares, Ruben R. ...
-
Varela, João C.
-
Neogi, Ujjwal
-
Ciftci, Sibel
-
Ashokkumar, Mani ...
-
Pinto, Ines Fern ...
-
visa fler...
-
Nilsson, Mats
-
Madaboosi, Naray ...
-
Russom, Aman, As ...
-
visa färre...
- Om ämnet
-
- NATURVETENSKAP
-
NATURVETENSKAP
-
och Biologi
-
och Biokemi och mole ...
-
- NATURVETENSKAP
-
NATURVETENSKAP
-
och Biologi
- Artiklar i publikationen
-
Biosensors & bio ...
- Av lärosätet
-
Kungliga Tekniska Högskolan
-
Stockholms universitet
-
Karolinska Institutet