Facile silicification of plastic surface for bioassays

Hong, Seonki; Park, Ki Soo; Weissleder, Ralph; Castro, Cesar M.; Lee, Hakho

doi:10.1039/c6cc09359k

Cited by 7 publications

(12 citation statements)

References 32 publications

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“…Thermoplastics have poor chemical and biological availability due to their intrinsic hydrophobicity and lack of nucleophilic functional groups for bioconjugation. To prepare our device for NA assays, we modified the device surface through a bioinspired silica coating process 22 . Silica is necessary in NA-based bioassays due to its intrinsic affinity to NA in the presence of chaotropic agents, but silica's hydrophilicity prevents its direct attachment/coating to hydrophobic thermoplastics.…”

Section: Resultsmentioning

confidence: 99%

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Rapid and simple single-chamber nucleic acid detection system prepared through nature-inspired surface engineering

et al. 2021

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Background: Nucleic acid (NA)-based diagnostics enable a rapid response to various diseases, but current techniques often require multiple labor-intensive steps, which is a major obstacle to successful translation to a clinical setting. Methods: We report on a surface-engineered single-chamber device for NA extraction and in situ amplification without sample transfer. Our system has two reaction sites: a NA extraction chamber whose surface is patterned with micropillars and a reaction chamber filled with reagents for in situ polymerase-based NA amplification. These two sites are integrated in a single microfluidic device; we applied plastic injection molding for cost-effective, mass-production of the designed device. The micropillars were chemically activated via a nature-inspired silica coating to possess a specific affinity to NA. Results: As a proof-of-concept, a colorimetric pH indicator was coupled to the on-chip analysis of NA for the rapid and convenient detection of pathogens. The NA enrichment efficiency was dependent on the lysate incubation time, as diffusion controls the NA contact with the engineered surface. We could detect down to 1×10 3 CFU by the naked eye within one hour of the total assay time. Conclusion: We anticipate that the surface engineering technique for NA enrichment could be easily integrated as a part of various types of microfluidic chips for rapid and convenient nucleic acid-based diagnostics.

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Section: Resultsmentioning

confidence: 99%

“…The process of pDA-mediated silica coating on the chip was modified slightly from a previous study 22 . Briefly, the top and bottom plates of a polycarbonate chip were incubated in 2 mg/mL dopamine hydrochloride (Sigma Aldrich) in 1× PBS (Corning®) at pH 8.3 for 3 h at room temperature with gentle shaking.…”

Section: Methodsmentioning

confidence: 99%

Rapid and simple single-chamber nucleic acid detection system prepared through nature-inspired surface engineering

et al. 2021

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“…The DNA extraction device is built on a bead-loaded disposable filter to directly isolate DNA from cell lysates (Figure S1) 19. We used poly(methyl methacrylate) microbeads (PMMA, 14.7 µm in diameter) coated with a silica layer via polydopamine to capture negatively charged DNA on the silica surface.…”

Section: Resultsmentioning

confidence: 99%

“…We used poly(methyl-methacrylate) microbeads (PMMA, 14.7 µm in diameter, Bangs Laboratories) with silica coating19 for DNA isolation. The beads (20 mg/mL) were incubated in PBS at pH 8.3, containing 2 mg/mL of dopamine (Sigma Aldrich) for 1 h, after washing with distilled water.…”

Section: Methodsmentioning

confidence: 99%

Point-of-care cervical cancer screening using deep learning-based microholography

Pathania¹,

Landeros²,

Rohrer³

et al. 2019

Theranostics

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Most deaths (80%) from cervical cancer occur in regions lacking adequate screening infrastructures or ready access to them. In contrast, most developed countries now embrace human papillomavirus (HPV) analyses as standalone screening; this transition threatens to further widen the resource gap.Methods: We describe the development of a DNA-focused digital microholography platform for point-of-care HPV screening, with automated readouts driven by customized deep-learning algorithms. In the presence of high-risk HPV 16 or 18 DNA, microbeads were designed to bind the DNA targets and form microbead dimers. The resulting holographic signature of the microbeads was recorded and analyzed.Results: The HPV DNA assay showed excellent sensitivity (down to a single cell) and specificity (100% concordance) in detecting HPV 16 and 18 DNA from cell lines. Our deep learning approach was 120-folder faster than the traditional reconstruction method and completed the analysis in < 2 min using a single CPU. In a blinded clinical study using patient cervical brushings, we successfully benchmarked our platform's performance to an FDA-approved HPV assay.Conclusions: Reliable and decentralized HPV testing will facilitate cataloguing the high-risk HPV landscape in underserved populations, revealing HPV coverage gaps in existing vaccination strategies and informing future iterations.

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“…Incorporating RNA extraction into the assay system would realize a true “sample-in and answer-out” test. One promising strategy is to integrate solid-phase extraction (e.g., silica beads) ( Hong et al, 2017 ) in a detachable cartridge. Increasing the assay throughput is also necessary, particularly for COVID-19 diagnostics, to detect all three targets (N1, N2, RPP30) at the same time.…”

Section: Discussionmentioning

confidence: 99%