scholarly journals Glycerol Reduces Cross Hybridization on Nitrocellulose Membrane

2020 ◽  
Vol 38 (3) ◽  
pp. 199
Author(s):  
Narendra Yoga Hendarta ◽  
Abu Tholib Aman ◽  
Asmarani Kusumawati ◽  
Tri Wibawa
Keyword(s):  
Nucleic Acid ◽  
Point Of Care ◽  
Gc Content ◽  
Lateral Flow ◽  
Hybridization Signal ◽  
Nitrocellulose Membrane ◽  
High Concentration ◽  
Cross Hybridization ◽  
High Stringency ◽  
Stringency Condition

Lateral flow assay (LFD) based nucleic acid lateral flow (NALF)  method has been developed recently. The method met point of care testing (POCT) as simple and rapid procedures, less equipment, and can be performance by less skilled technician. NALF based on nucleic acid hybridizationis  more economical then immunochromatography assay which use antibody-antigen recognition. Cross hybridization has issued while used to differentiate organism with high GC content and high homology as high similarity genome. Some techniques has applied to give high stringency condition avoid cross hybridization reaction but need more procedure to apply. We found glycerol applied to buffer assay could reduce cross hybridization on nitrocellulose membrane. The study used 2 kinds of high stringency buffer as PBS and SSC bases and high concentration of ssDNA amplicon as sample. Without glycerol ingredient gave cross hybridization signal on test line. But used glycerol could reduce those even omitted with PBS based buffer assay. Beside those, glycerol could significantly increased hybridization signal in SSC based buffer assay (p<0.05).

Gold-Aptamer-Nanoconstructs Engineered to Detect Conserved Enteroviral Nucleic Acid Sequences

2019 ◽  
Author(s):  
Veeren Chauhan ◽  
Mohamed M Elsutohy ◽  
C Patrick McClure ◽  
Will Irving ◽  
Neil Roddis ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
In Silico ◽  
Point Of Care ◽  
Lateral Flow ◽  
Nucleic Acid Sequence ◽  
In Silico Screening ◽  
Lateral Flow Assays ◽  
Life Threatening ◽  
Software And Hardware ◽  
Nucleic Acid Sequences

<p>Enteroviruses are a ubiquitous mammalian pathogen that can produce mild to life-threatening disease. Bearing this in mind, we have developed a rapid, accurate and economical point-of-care biosensor that can detect a nucleic acid sequences conserved amongst 96% of all known enteroviruses. The biosensor harnesses the physicochemical properties of gold nanoparticles and aptamers to provide colourimetric, spectroscopic and lateral flow-based identification of an exclusive enteroviral RNA sequence (23 bases), which was identified through in silico screening. Aptamers were designed to demonstrate specific complementarity towards the target enteroviral RNA to produce aggregated gold-aptamer nanoconstructs. Conserved target enteroviral nucleic acid sequence (≥ 1x10<sup>-7</sup> M, ≥1.4×10<sup>-14</sup> g/mL), initiates gold-aptamer-nanoconstructs disaggregation and a signal transduction mechanism, producing a colourimetric and spectroscopic blueshift (544 nm (purple) > 524 nm (red)). Furthermore, lateral-flow-assays that utilise gold-aptamer-nanoconstructs were unaffected by contaminating human genomic DNA, demonstrated rapid detection of conserved target enteroviral nucleic acid sequence (< 60 s) and could be interpreted with a bespoke software and hardware electronic interface. We anticipate our methodology will translate in-silico screening of nucleic acid databases to a tangible enteroviral desktop detector, which could be readily translated to related organisms. This will pave-the-way forward in the clinical evaluation of disease and complement existing strategies at overcoming antimicrobial resistance.</p>

scholarly journals Eliminating viscosity bias in lateral flow tests

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Daniel M. Kainz ◽  
Bastian J. Breiner ◽  
Susanna M. Früh ◽  
Tobias Hutzenlaub ◽  
Roland Zengerle ◽  
...  
Keyword(s):  
Point Of Care ◽  
Lateral Flow ◽  
Nitrocellulose Membrane ◽  
Widespread Application ◽  
Significant Challenge ◽  
Microfluidic Flow ◽  
Flow Test ◽  
Lateral Flow Test ◽  
Lateral Flow Tests ◽  
Viscosity Dependence

AbstractDespite the widespread application of point-of-care lateral flow tests, the viscosity dependence of these assay results remains a significant challenge. Here, we employ centrifugal microfluidic flow control through the nitrocellulose membrane of the strip to eliminate the viscosity bias. The key feature is the balancing of the sample flow into the cassette of the lateral flow test with the air flow out of the cassette. A viscosity-independent flow rate of 3.01 ± 0.18 µl/min (±6%) is demonstrated for samples with viscosities ranging from 1.1 mPas to 24 mPas, a factor greater than 20. In a model human IgG lateral flow assay, signal-intensity shifts caused by varying the sample viscosity from 1.1 mPas to 2.3 mPas could be reduced by more than 84%.

Gold-Aptamer-Nanoconstructs Engineered to Detect Conserved Enteroviral Nucleic Acid Sequences

2019 ◽  
Author(s):  
Veeren Chauhan ◽  
Mohamed M Elsutohy ◽  
C Patrick McClure ◽  
Will Irving ◽  
Neil Roddis ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
In Silico ◽  
Point Of Care ◽  
Lateral Flow ◽  
Nucleic Acid Sequence ◽  
In Silico Screening ◽  
Lateral Flow Assays ◽  
Life Threatening ◽  
Software And Hardware ◽  
Nucleic Acid Sequences

<p>Enteroviruses are a ubiquitous mammalian pathogen that can produce mild to life-threatening disease. Bearing this in mind, we have developed a rapid, accurate and economical point-of-care biosensor that can detect a nucleic acid sequences conserved amongst 96% of all known enteroviruses. The biosensor harnesses the physicochemical properties of gold nanoparticles and aptamers to provide colourimetric, spectroscopic and lateral flow-based identification of an exclusive enteroviral RNA sequence (23 bases), which was identified through in silico screening. Aptamers were designed to demonstrate specific complementarity towards the target enteroviral RNA to produce aggregated gold-aptamer nanoconstructs. Conserved target enteroviral nucleic acid sequence (≥ 1x10<sup>-7</sup> M, ≥1.4×10<sup>-14</sup> g/mL), initiates gold-aptamer-nanoconstructs disaggregation and a signal transduction mechanism, producing a colourimetric and spectroscopic blueshift (544 nm (purple) > 524 nm (red)). Furthermore, lateral-flow-assays that utilise gold-aptamer-nanoconstructs were unaffected by contaminating human genomic DNA, demonstrated rapid detection of conserved target enteroviral nucleic acid sequence (< 60 s) and could be interpreted with a bespoke software and hardware electronic interface. We anticipate our methodology will translate in-silico screening of nucleic acid databases to a tangible enteroviral desktop detector, which could be readily translated to related organisms. This will pave-the-way forward in the clinical evaluation of disease and complement existing strategies at overcoming antimicrobial resistance.</p>

Polydimethylsiloxane-Paper Hybrid Lateral Flow Assay for Highly Sensitive Point-of-Care Nucleic Acid Testing

2016 ◽  
Vol 88 (12) ◽  
pp. 6254-6264 ◽  
Author(s):  
Jane Ru Choi ◽  
Zhi Liu ◽  
Jie Hu ◽  
Ruihua Tang ◽  
Yan Gong ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Point Of Care ◽  
Lateral Flow ◽  
Lateral Flow Assay ◽  
Nucleic Acid Testing ◽  
Highly Sensitive

Electrospin-coating of nitrocellulose membrane enhances sensitivity in nucleic acid-based lateral flow assay

2018 ◽  
Vol 1009 ◽  
pp. 81-88 ◽  
Author(s):  
Chee-Hong Takahiro Yew ◽  
Pedram Azari ◽  
Jane Ru Choi ◽  
Fei Li ◽  
Belinda Pingguan-Murphy
Keyword(s):  
Nucleic Acid ◽  
Lateral Flow ◽  
Lateral Flow Assay ◽  
Nitrocellulose Membrane

scholarly journals Gold–Oligonucleotide Nanoconstructs Engineered to Detect Conserved Enteroviral Nucleic Acid Sequences

Biosensors ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 238
Author(s):  
Veeren M. Chauhan ◽  
Mohamed M. Elsutohy ◽  
C. Patrick McClure ◽  
William L. Irving ◽  
Neil Roddis ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
In Silico ◽  
Point Of Care ◽  
Lateral Flow ◽  
Nucleic Acid Sequence ◽  
In Silico Screening ◽  
Lateral Flow Assays ◽  
Life Threatening ◽  
Software And Hardware ◽  
Nucleic Acid Sequences

Enteroviruses are ubiquitous mammalian pathogens that can produce mild to life-threatening disease. We developed a multimodal, rapid, accurate and economical point-of-care biosensor that can detect nucleic acid sequences conserved amongst 96% of all known enteroviruses. The biosensor harnesses the physicochemical properties of gold nanoparticles and oligonucleotides to provide colourimetric, spectroscopic and lateral flow-based identification of an exclusive enteroviral nucleic acid sequence (23 bases), which was identified through in silico screening. Oligonucleotides were designed to demonstrate specific complementarity towards the target enteroviral nucleic acid to produce aggregated gold–oligonucleotide nanoconstructs. The conserved target enteroviral nucleic acid sequence (≥1 × 10−7 M, ≥1.4 × 10−14 g/mL) initiates gold–oligonucleotide nanoconstruct disaggregation and a signal transduction mechanism, producing a colourimetric and spectroscopic blueshift (544 nm (purple) > 524 nm (red)). Furthermore, lateral-flow assays that utilise gold–oligonucleotide nanoconstructs were unaffected by contaminating human genomic DNA, demonstrated rapid detection of conserved target enteroviral nucleic acid sequence (<60 s), and could be interpreted with a bespoke software and hardware electronic interface. We anticipate that our methodology will translate in silico screening of nucleic acid databases to a tangible enteroviral desktop detector, which could be readily translated to related organisms. This will pave the way forward in the clinical evaluation of disease and complement existing strategies to overcome antimicrobial resistance.

Point-of-care and visual detection of P. aeruginosa and its toxin genes by multiple LAMP and lateral flow nucleic acid biosensor

2016 ◽  
Vol 81 ◽  
pp. 317-323 ◽  
Author(s):  
Yuting Chen ◽  
Nan Cheng ◽  
Yuancong Xu ◽  
Kunlun Huang ◽  
Yunbo Luo ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Visual Detection ◽  
Point Of Care ◽  
Lateral Flow ◽  
Toxin Genes

scholarly journals Development of nucleic acid aptamer-based lateral flow assays: A robust platform for cost-effective point-of-care diagnosis

Theranostics ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 5174-5196
Author(s):  
Tao Wang ◽  
Lanmei Chen ◽  
Arpitha Chikkanna ◽  
Suxiang Chen ◽  
Isabell Brusius ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Point Of Care ◽  
Cost Effective ◽  
Lateral Flow ◽  
Lateral Flow Assays
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