1. 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 http://www.who.int/bulletin/volumes/90/12/BLT-12-102780-table-T1.html http://www.finddiagnostics.org/ http://developeconomies.com/ http://www.popularmechanics.com http://jaffeweb.ucsd.edu/node/81 http://www.finddiagnostics.org/ http://www.forbes.com/sites/gcaptain/2011/10/04/the- ocean-network

2. 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, 2008. c

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