Fully automated sample to result SIMPLE RPA microfluidic chip towards in ovo sexing application DOI Creative Commons
Simão Santos, Celine Wegsteen, Dries Vloemans

et al.

Deleted Journal, Journal Year: 2024, Volume and Issue: 1(1)

Published: Dec. 26, 2024

Several European countries have implemented legislations to eliminate day-old male chicks killing. Although embryo sexing (in ovo sexing) is the most promising alternative, no current solution can handle all egg colors with >98% accuracy, low cost and minimal disturbance before day 13 of incubation processing >20,000 eggs/hour. Recombinase polymerase amplification (RPA) presents a alternative PCR that be integrated into microfluidic platforms. In this work, we developed fully autonomous cartridge (SIMPLE-RPA chip) for female chick-specific synthetic HINTW gene detection in 30 min at 37.7 °C inside an incubator. We first optimized off-chip RPA, allowing highly sensitive DNA (1.6 × 10–5 ng/µL). The SIMPLE-RPA chip was automate RPA bioassay on-chip, reducing user errors, contamination risks maintaining LOD while offering price, small footprint, upscaling compatibility, easy transfer other point-of-care applications.

Language: Английский

Point-of-need diagnostics in a post-Covid world: an opportunity for paper-based microfluidics to serve during syndemics DOI Creative Commons
Maria‐Nefeli Tsaloglou, Dionysios C. Christodouleas,

Jonathan Milette

et al.

Lab on a Chip, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

A post-pandemic perspective on paper-based microfluidics by George Whitesides and collaborators. Sketch generated artificial intelligence using an original photograph courtesy of Volker Steger (Science Photo Library).

Language: Английский

Citations

0
Detection of Vibrio parahaemolyticus by one-pot LAMP-CRISPR/Cas12b combined with immunomagnetic beads DOI
Shi-Qi Wang,

Yongheng Gao,

Yiqin Miao

et al.

Food Control, Journal Year: 2025, Volume and Issue: unknown, P. 111181 - 111181

Published: Jan. 1, 2025

Language: Английский

Citations

0
Structural basis for intrinsic strand displacement activity of mitochondrial DNA polymerase DOI Creative Commons
Ashok R. Nayak, V. E. Sokolova, Sirelin Sillamaa

et al.

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: March 11, 2025

Members of the Pol A family DNA polymerases, found across all domains life, utilize various strategies for strand separation during replication. In higher eukaryotes, mitochondrial polymerase γ relies on replicative helicase TWINKLE, whereas yeast ortholog, Mip1, can unwind independently. Using Mip1 as a model, we present series high-resolution cryo-EM structures that capture process displacement. Our data reveal previously unidentified structural elements facilitate unwinding downstream duplex. Yeast cells harboring variants defective in displacement exhibit impaired oxidative phosphorylation and loss mtDNA, corroborating observations. This study provides molecular basis intrinsic activity illuminates distinct mechanisms utilized by polymerases. Mitochondrial polymerases use different Here, authors identifying key enable Mutations these impair function maintenance.

Language: Английский

Citations

0
Fully automated sample-to-result SIMPLE-RPA microfluidic chip: towards in ovo sexing application DOI
Simão Santos, Celine Wegsteen, Dries Vloemans

et al.

Research Square (Research Square), Journal Year: 2024, Volume and Issue: unknown

Published: Sept. 24, 2024

Abstract Several European countries have implemented new legislations to eliminate the killing of day-old male chicks, pushing laying hen industry find animal-friendly solutions. Although embryo sexing during incubation (in ovo sexing) is highly promising, no current solution meets requirements handling all egg colors with >98 % accuracy, low cost and minimal disturbance, while allowing before day 13 processing > 20 000 eggs/hour. Recombinase polymerase amplification (RPA) presents a promising alternative PCR as sensitive isothermal technique be integrated into microfluidic platform. In this work we developed fully autonomous cartridge (SIMPLE-RPA chip) for chick female-specific synthetic HINTW gene detection in 30 minutes at 37.7 °C inside an incubator. We first optimized off-chip RPA, DNA (1.6 x 10-5 ng/µL), matching expected concentrations allantoic fluid samples. Then, SIMPLE-RPA chip was accommodate automate RPA bioassay on-chip, requiring only sample introduction finger press activation reduce user errors contamination risks. Importantly, maintained LOD achieved off-chip. The presented inexpensive, has small footprint, compatible upscaling, can easily transferred other point-of-care applications.

Language: Английский

Citations

0
Fully automated sample to result SIMPLE RPA microfluidic chip towards in ovo sexing application DOI Creative Commons
Simão Santos, Celine Wegsteen, Dries Vloemans

et al.

Deleted Journal, Journal Year: 2024, Volume and Issue: 1(1)

Published: Dec. 26, 2024

Several European countries have implemented legislations to eliminate day-old male chicks killing. Although embryo sexing (in ovo sexing) is the most promising alternative, no current solution can handle all egg colors with >98% accuracy, low cost and minimal disturbance before day 13 of incubation processing >20,000 eggs/hour. Recombinase polymerase amplification (RPA) presents a alternative PCR that be integrated into microfluidic platforms. In this work, we developed fully autonomous cartridge (SIMPLE-RPA chip) for female chick-specific synthetic HINTW gene detection in 30 min at 37.7 °C inside an incubator. We first optimized off-chip RPA, allowing highly sensitive DNA (1.6 × 10–5 ng/µL). The SIMPLE-RPA chip was automate RPA bioassay on-chip, reducing user errors, contamination risks maintaining LOD while offering price, small footprint, upscaling compatibility, easy transfer other point-of-care applications.

Language: Английский

Citations

0