Passively driven microfluidic device with simple operation in the development of nanolitre droplet assay in nucleic acid detection

Lin, Pei Heng; Li, Bor Ran

doi:10.1038/s41598-021-00470-9

Cited by 17 publications

(6 citation statements)

References 50 publications

(38 reference statements)

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“…There was a visible increase in the concatemeric signal intensity as the Ct value decreased, corresponding to the higher viral load in the sample. The gel electrophoresis result showed that RT-LAMP could amplify swab samples with the highest (33) and lowest (18.55) Ct values within 20 min of processing time at 65 • C.…”

Section: Optimisation Of Processing Times For Swab Sample Amplificati...mentioning

confidence: 99%

Detection of Reverse Transcriptase LAMP-Amplified Nucleic Acid from Oropharyngeal Viral Swab Samples Using Biotinylated DNA Probes through a Lateral Flow Assay

Agarwal,

Hamidizadeh,

Bier

2023

Biosensors

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This study focuses on three key aspects: (a) crude throat swab samples in a viral transport medium (VTM) as templates for RT-LAMP reactions; (b) a biotinylated DNA probe with enhanced specificity for LFA readouts; and (c) a digital semi-quantification of LFA readouts. Throat swab samples from SARS-CoV-2 positive and negative patients were used in their crude (no cleaning or pre-treatment) forms for the RT-LAMP reaction. The samples were heat-inactivated but not treated for any kind of nucleic acid extraction or purification. The RT-LAMP (20 min processing time) product was read out by an LFA approach using two labels: FITC and biotin. FITC was enzymatically incorporated into the RT-LAMP amplicon with the LF-LAMP primer, and biotin was introduced using biotinylated DNA probes, specifically for the amplicon region after RT-LAMP amplification. This assay setup with biotinylated DNA probe-based LFA readouts of the RT-LAMP amplicon was 98.11% sensitive and 96.15% specific. The LFA result was further analysed by a smartphone-based IVD device, wherein the T-line intensity was recorded. The LFA T-line intensity was then correlated with the qRT-PCR Ct value of the positive swab samples. A digital semi-quantification of RT-LAMP-LFA was reported with a correlation coefficient of R2 = 0.702. The overall RT-LAMP-LFA assay time was recorded to be 35 min with a LoD of three RNA copies/µL (Ct-33). With these three advancements, the nucleic acid testing-point of care technique (NAT-POCT) is exemplified as a versatile biosensor platform with great potential and applicability for the detection of pathogens without the need for sample storage, transportation, or pre-processing.

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Section: Optimisation Of Processing Times For Swab Sample Amplificati...mentioning

confidence: 99%

Detection of Reverse Transcriptase LAMP-Amplified Nucleic Acid from Oropharyngeal Viral Swab Samples Using Biotinylated DNA Probes through a Lateral Flow Assay

Agarwal,

Hamidizadeh,

Bier

2023

Biosensors

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“…Among all water contamination, heavy metal ions such as Pb(II), Cd(II), Zn(II), Ni(II) and Hg(II) are highly toxic and non-biodegradable, potentially causing serious health problems in animals. and humans (P. H. Lin & Li, 2021;Shari et al, 2022).…”

Section: Introductionmentioning

confidence: 99%

Paper-based device (PAD) for simultaneous determination of Hg(II) and Pb(II) in water samples from São Francisco River, Brazil

Silva,

Soares,

Júnior

et al. 2024

Preprint

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Hg(II) and Pb(II) are considered potentially toxic metals (PTMs) that can be found in several ecosystems, including water systems such as lakes, rivers and groundwaters. These species are classified as human carcinogens and non-biodegradable pollutants, leading to neurological and nutritional disorders, as well as chronic diseases. Due to its relevance, several methods have been developed to determine the concentration of these PTMs; however, they require costly reagents, laborious operation and complex equipment. Herein, we present a simple-to-use and low-cost paper-based device (PAD) for Hg(II) and Pb(II) determination in natural water samples based on the colorimetric reaction between these PMTs and the dithizone colorimetric probe. The color intensity was employed as an analytical signal, through the color channels of the RGB scale. Under optimized conditions, a linear correlation of r = 0.989 and r = 0.991 and linear range of 1–20 µg/mL and 5–30 µg/mL were obtained for Hg(II) and Pb(II), respectively. After analytical optimization, the method was employed for PMTs determination using real samples from São Francisco River, Brazil. Thus, this method proved to be appropriate for Hg(II) and Pb(II) determination in natural water samples and is a easy, low-cost and straightforward alternative for the simultaneous determination of Hg(II) and Pb(II).

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“…Microfluidics is multidisciplinary for precise regulation and manipulation in geometrically confined small-scale fluids (22). Microfluidics has practical applications, such as cell size sorting, particle formation, particle trapping, and gradient generation, and is widely applied in the fields of organs-on-chip, cell culture, drug delivery, biochemistry, and biotechnology (23)(24)(25). Microfluidics offers great opportunities for integration with wearable sensors that complete the sample-to-answer solution (26).…”

Section: Introductionmentioning

confidence: 99%

Wearable Microfluidics for Continuous Assay

Lin

Nien

2023

Annual Rev. Anal. Chem.

Self Cite

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The development of wearable devices provides approaches for the realization of self-health care. Easily carried wearable devices allow individual health monitoring at any place whenever necessary. There are various interesting monitoring targets, including body motion, organ pressure, and biomarkers. An efficient use of space in one small device is a promising resolution to increase the functions of wearable devices. Through integration of a microfluidic system into wearable devices, embedding complicated structures in one design becomes possible and can enable multifunction analyses within a limited device volume. This article reviews the reported microfluidic wearable devices, introduces applications to different biofluids, discusses characteristics of the design strategies and sensing principles, and highlights the attractive configurations of each device. This review seeks to provide a detailed summary of recent advanced microfluidic wearable devices. The overview of advanced key components is the basis for the development of future microfluidic wearable devices. Expected final online publication date for the Annual Review of Analytical Chemistry, Volume 16 is June 2023. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

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Passively driven microfluidic device with simple operation in the development of nanolitre droplet assay in nucleic acid detection

Cited by 17 publications

References 50 publications

Detection of Reverse Transcriptase LAMP-Amplified Nucleic Acid from Oropharyngeal Viral Swab Samples Using Biotinylated DNA Probes through a Lateral Flow Assay

Detection of Reverse Transcriptase LAMP-Amplified Nucleic Acid from Oropharyngeal Viral Swab Samples Using Biotinylated DNA Probes through a Lateral Flow Assay

Paper-based device (PAD) for simultaneous determination of Hg(II) and Pb(II) in water samples from São Francisco River, Brazil

Wearable Microfluidics for Continuous Assay

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