Ultrasensitive nucleic acid biosensor based on enzyme–gold nanoparticle dual label and lateral flow strip biosensor

He, Yuqing; Zhang, Sanquan; Zhang, Xibao; Baloda, Meenu; Gurung, Anant S.; Xu, Hui; Zhang, Xueji; Liu, Guodong

doi:10.1016/j.bios.2010.08.079

Cited by 196 publications

(131 citation statements)

References 22 publications

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“…Previous strategies to improve lateral flow immunoassay performance have included adopting a two-step assay for enzymatic colorimetric signal generation or prestabilization of reactants. (2,15,16) Our results showed a dot intensity increase similar to that obtained by Parolo et al, (2) who demonstrated a 3-or 4-fold intensity increase by adopting a two-step assay using an enzymatic colorimetric signal, although their system also increased the analytical sensitivity by an order of magnitude. Changing the strip architecture resulted in a 2-or 3-fold change in signal intensity.…”

Section: Lateral Flow Immunoassay With Different Running Methodssupporting

confidence: 87%

Enhancing the Signal of Lateral Flow Immunoassays by Using Different Developing Methods

2015

Sensors and Materials

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Lateral flow immunoassay devices are potential biosensors for point-of-care diagnostics. However, these devices are limited by low analytical sensitivity when coupled with a visual colorimetric signal. Here, we analyzed key parameters related to nucleic acid lateral flow performance and related this to the analytical theory behind lateral flow functionality. In particular, we predicted a set of different physical running methods to optimize the assay signal intensity by ensuring higher binding of the signal molecules to the DNA. We found that a double-run method improved the assay signal intensity on average by approximately 50%, and a fully absorbed conjugate pad method came close to the double-run signal efficacy at high DNA concentrations. Our results demonstrate that even when the signal molecule is supplied in excess, a significant proportion of analytes bind to the detection antibody in a colorless complex. The lowest detection limits achieved were 0.1 picomole for detection using an anti-Cy5 antibody and 0.01 picomole for the detection using an anti-Texas Red antibody. Our double-run method improvement is generic, does not require additional reagents and equipment, reduces assay costs compared to the fully absorbed method, and is applicable to other lateral flow immunoassays.

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Section: Lateral Flow Immunoassay With Different Running Methodssupporting

confidence: 87%

Enhancing the Signal of Lateral Flow Immunoassays by Using Different Developing Methods

2015

Sensors and Materials

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“…25 However, the existing LFAs are associated with the limitation of poor sensitivity. 11,26 Several methods have been developed to enhance the sensitivity of LFAs, such as sample concentration, [27][28][29][30] thermal contrast, 31 fluidic control, [32][33][34] temperature-humidity technique, 35 probe-based signal enhancement, 23,36 enzyme-based signal amplification 37,38 and electrochemical devices-based enhancement. 39 However, these approaches require either external equipment, 35 high-cost reagents 23,37 or complicated fabrication with multistep procedure, 32 limiting their application for POC testing.…”

Section: Introductionmentioning

confidence: 99%

Improved LFIAs for highly sensitive detection of BNP at point-of-care

Gong¹,

Hu²,

Choi³

et al. 2017

IJN

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Heart failure (HF) has become a major cause of morbidity and mortality with a significant global economic burden. Although well-established clinical tests could provide early diagnosis, access to these tests is limited in developing countries, where a relatively higher incidence of HF is present. This has prompted an urgent need for developing a cost-effective, rapid and robust diagnostic tool for point-of-care (POC) detection of HF. Lateral flow immunoassay (LFIA) has found widespread applications in POC diagnostics. However, the low sensitivity of LFIA limits its ability to detect important HF biomarkers (e.g., brain natriuretic peptide [BNP]) that are normally present in low concentration in blood. To address this issue, we developed an improved LFIA by optimizing the gold nanoparticle (GNP)–antibody conjugate conditions (e.g., the conjugate pH and the amount of added antibody), the diameter of GNP and the concentration of antibody embedded on the test line and modifying the structure of test strip. Through these improvements, the proposed test strip enabled the detection of BNP down to 0.1 ng/mL within 10–15 min, presenting ~15-fold sensitivity enhancement over conventional lateral flow assay. We also successfully applied our LFIA in the analysis of BNP in human serum samples, highlighting its potential use for clinical assessment of HF. The developed LFIA for BNP could rapidly rule out HF with the naked eye, offering tremendous potential for POC test and personalized medicine.

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“…The lateral flow strip biosensor consisted of four components: sample application pad, conjugate pad, nitrocellulose membrane, and absorbent pad. 19 All components were mounted on a common backing layer (typically an inert plastic, e.g., polyester) using the Clamshell Laminator (Biodot; Irvin, CA).…”

Section: Preparation Of Gold-nanoparticle Coated Silica Nanorod (Gnpsmentioning

confidence: 99%

Gold Nanoparticle Coated Silica Nanorods for Sensitive Visual Detection of microRNA on a Lateral Flow Strip Biosensor

Takalkar

Chen

et al. 2016

ANAL. SCI.

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We present a rapid and highly sensitive approach for visual detection of microRNA (miRNA) using a gold nanoparticles coated silica nanorod label and lateral flow strip biosensor. Gold nanoparticles were decorated on the silica nanorod surface by a seeding and growth procedure. A single strand DNA probe was immobilized on the gold nanoparticles-silica nanorod surface by a self-assembling process, and the formed DNA-gold nanoparticles-silica nanorod conjugate was used to construct the lateral flow nucleic acid biosensor for detecting miRNA. The captured gold nanoparticles-silica nanorods by sandwich-type hybridization reactions (DNA-RNA-DNA) on the test zone of the lateral flow nucleic acid biosensor produced the characteristic color bands, enabling visual detection of miRNA. After systematic optimization, the new lateral flow nucleic acid biosensor was capable of detecting 10 pM of the miRNA target without instrumentation, which is six times lower than that obtained with the gold nanoparticle-based lateral flow nucleic acid biosensor. The gold nanoparticles coated silica nanorod thus provides a new and sensitive nanolabel for visual detection of biological molecules on the lateral flow biosensor.

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Ultrasensitive nucleic acid biosensor based on enzyme–gold nanoparticle dual label and lateral flow strip biosensor

Cited by 196 publications

References 22 publications

Enhancing the Signal of Lateral Flow Immunoassays by Using Different Developing Methods

Enhancing the Signal of Lateral Flow Immunoassays by Using Different Developing Methods

Improved LFIAs for highly sensitive detection of BNP at point-of-care

Gold Nanoparticle Coated Silica Nanorods for Sensitive Visual Detection of microRNA on a Lateral Flow Strip Biosensor

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