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Circle-to-circle am...
Circle-to-circle amplification coupled with microfluidic affinity chromatography enrichment for in vitro molecular diagnostics of Zika fever and analysis of anti-flaviviral drug efficacy
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- Soares, Ruben R. G. (author)
- 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)
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- Pettke, Aleksandra (author)
- Karolinska Inst, Dept Oncol Pathol, Sci Life Lab, Solna, Sweden.
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- Robles-Remacho, Agustin (author)
- Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab),Stockholm Univ, Dept Biochem & Biophys, Sci Life Lab, Solna, Sweden.
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- Zeebaree, Sahar (author)
- Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab),Stockholm Univ, Dept Biochem & Biophys, Sci Life Lab, Solna, Sweden.
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- Ciftci, Sibel (author)
- Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab),Stockholm Univ, Dept Biochem & Biophys, Sci Life Lab, Solna, Sweden.
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- Tampere, Marianna (author)
- Karolinska Institutet
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- Russom, Aman, Prof. 1976- (author)
- KTH,Science for Life Laboratory, SciLifeLab,Nanobioteknologi,Karolinska Inst, AIMES Ctr Adv Integrated Med & Engn Sci, Stockholm, Sweden.;KTH Royal Inst Technol, Stockholm, Sweden.
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- Puumalainen, Marjo-Riitta (author)
- Karolinska Inst, Dept Oncol Pathol, Sci Life Lab, Solna, Sweden.
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- Nilsson, Mats (author)
- Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab),Stockholm Univ, Dept Biochem & Biophys, Sci Life Lab, Solna, Sweden.
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- Madaboosi, Narayanan (author)
- Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab),Stockholm Univ, Dept Biochem & Biophys, Sci Life Lab, Solna, Sweden.;Indian Inst Technol Madras, Chennai, Tamil Nadu, India.
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(creator_code:org_t)
- Elsevier BV, 2021
- 2021
- English.
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In: Sensors and actuators. B, Chemical. - : Elsevier BV. - 0925-4005 .- 1873-3077. ; 336
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Abstract
Subject headings
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- Sensitive viral diagnostic methods are increasingly in demand to tackle emerging epidemics. The Zika virus (ZIKV) is particularly relevant in tropical resource limited settings (RLS) and is associated with intermittent epidemics such as the recent 2016 ZIKV outbreak in South America, wherein Zika fever was classified by WHO as a public health emergency of international concern. Thus, there is an urgent need for widespread Zika fever diagnostics and efficient drug therapies. ZIKV diagnostics are typically performed using RT-qPCR in centralized laboratories. While extremely sensitive, RT-qPCR requires rapid heating-cooling cycles, combined with continuous fluorescence measurements to allow quantification, implying high costs and limiting availability of molecular diagnostics in RLS. Here, we report isothermal amplification of ZIKV cDNA using padlock probes followed by two rounds of Rolling Circle Amplification (RCA), termed as circle-to-circle amplification (C2CA), combined with a microfluidic affinity chromatography enrichment (mu ACE) platform. This platform allowed the detection of <17 vRNA copies per reaction mixture, equivalent to similar to 3 aM, showed a positive correlation with RT-qPCR in both average (r = 0.80) and discrete (r = 0.95) signal modes, and was validated for drug efficiency tests using in vitro infected peripheral blood mononuclear cells from 3 healthy donors. This performance shows significant promise towards highly sensitive, albeit simple and cost-effective point-of-care viral diagnostics.
Subject headings
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Medicinsk genetik (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Medical Genetics (hsv//eng)
- NATURVETENSKAP -- Kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences (hsv//eng)
Keyword
- Rolling circle amplification
- Zika virus
- Circle-to-circle amplification
- Padlock probes
- Diagnostics
- Microfluidics
- Streptavidin beads
Publication and Content Type
- ref (subject category)
- art (subject category)
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Soares, Ruben R. ...
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Pettke, Aleksand ...
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Robles-Remacho, ...
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Zeebaree, Sahar
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Ciftci, Sibel
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Tampere, Mariann ...
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Russom, Aman, Pr ...
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Puumalainen, Mar ...
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Nilsson, Mats
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Madaboosi, Naray ...
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- About the subject
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
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and Biochemistry and ...
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- MEDICAL AND HEALTH SCIENCES
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MEDICAL AND HEAL ...
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and Basic Medicine
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and Medical Genetics
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Chemical Science ...
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Sensors and actu ...
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Royal Institute of Technology
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Stockholm University
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Karolinska Institutet