ASSURED criteria (WHO) for the evaluation of point-of-care devices CharacteristicExample target specification Affordable Less than US$ 500 per machine,

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Research Techniques Made Simple: Polymerase Chain Reaction

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ASSURED criteria (WHO) for the evaluation of point-of-care devices CharacteristicExample target specification Affordable Less than US$ 500 per machine, less than US$ 10 per test Sensitivity, Specificity Lower limit of detection: 500 HIV RNA copies per mL, 350 CD4+ T-cells per μL User-friendly1–2 days of training, easy to use Rapid and robust 1 year at room temperature, high throughput Equipment-free Compact, battery powered, on-site data analysis, easy disposal, easy sample handling, no cold chain DeliverablePortable, hand-held ocean-network

The LAMP reaction Features:  Isothermal (60-65 °C)  No heat denature required  High gain: ~10 9  PCR-like sensitivity, selectivity  Robustness to many PCR inhibitors  ‘One-pot’ DNA or RNA Tomita, et al. “Loop-mediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products”, Nature Protocols, Vol. 3 No. 5, pg. 877, c

- +++ t=0, room temperature t=1 h, reaction (65 o C ) ~20 Background of 100 ng/µl Salmon sperm DNA