Point-of-care isothermal nucleic acid amplification tests: progress and bottlenecks for extraction-free sample collection and preparation

Wilkinson, Alexis F.; Barra, Maria J.; Novak, Emilie N.; Bond, Meaghan; Richards-Kortum, Rebecca

doi:10.1080/14737159.2024.2375233

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2024

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Cited by 2 publications

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Fully automated sample to result SIMPLE RPA microfluidic chip towards in ovo sexing application

Monteiro Belo dos Santos,

Wegsteen,

Vloemans

et al. 2024

npj Biosensing

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Fully automated sample to result SIMPLE RPA microfluidic chip towards in ovo sexing application

Monteiro Belo dos Santos,

Wegsteen,

Vloemans

et al. 2024

npj Biosensing

View full text Add to dashboard Cite

Fully automated sample-to-result SIMPLE-RPA microfluidic chip: towards in ovo sexing application

Santos,

Wegsteen,

Vloemans

et al. 2024

Preprint

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

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Point-of-care isothermal nucleic acid amplification tests: progress and bottlenecks for extraction-free sample collection and preparation

Cited by 2 publications

References 87 publications

Fully automated sample to result SIMPLE RPA microfluidic chip towards in ovo sexing application

Fully automated sample to result SIMPLE RPA microfluidic chip towards in ovo sexing application

Fully automated sample-to-result SIMPLE-RPA microfluidic chip: towards in ovo sexing application

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