http www finddiagnostics org http jaffeweb ucsd edunode81
http: //www. finddiagnostics. org/ http: //jaffeweb. ucsd. edu/node/81 http: //www. finddiagnostics. org/ ASSURED criteria (WHO) for the evaluation of point-of-care devices Characteristic Affordable Sensitivity, Specificity User-friendly Rapid and robust Equipment-free Deliverable http: //developeconomies. com/ Example target specification Less than US$ 500 per machine, less than US$ 10 per test Lower limit of detection: 500 HIV RNA copies per m. L, 350 CD 4+ T-cells per μL http: //www. popularmechanics. com 1– 2 days of training, easy to use < 30 minutes for diagnosis, < 1. 5 hours for HIV load monitoring, minimal consumables (i. e. pipettes), shelf life > 1 year at room temperature, high throughput Compact, battery powered, on-site data analysis, easy disposal, easy sample handling, no cold chain Portable, hand-held http: //www. who. int/bulletin/volumes/90/12/BLT-12 -102780 -table-T 1. html http: //www. forbes. com/sites/gcaptain/2011/10/04/theocean-network
The LAMP reaction Features: § Isothermal (60 -65 °C) § No heat denature required § High gain: ~109 § PCR-like sensitivity, selectivity c § 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.
Ct (hours) t=0, room temperature 2 LAMP 1. 75 ASU ~20 1. 5 - 1. 25 1 + + t=1 h, reaction (65 o. C ) 0. 75 0. 5 0 2 4 6 Template copies per 10 ml (log 10) Background of 100 ng/µl Salmon sperm DNA 8 - + +
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