01996nas a2200205   4500008004100000022001400041245014400055210006900199260001300268300001400281490000700295520132200302100001601624700001801640700002301658700001301681700001801694700002301712856005501735       2013                        ENG d  a1001-653800aMolecular diagnostics in a teacup: Non-Instrumented Nucleic Acid Amplification (NINA) for rapid, low cost detection of Salmonella enterica.0 aMolecular diagnostics in a teacup NonInstrumented Nucleic Acid A  c2013 Apr  a1162-11680 v583 a<p>We report on the use of a novel non-instrumented platform to enable a Loop Mediated isothermal Amplification (LAMP) based assay for Salmonella enterica. Heat energy is provided by addition of a small amount (<150 g) of boiling water, and the reaction temperature is regulated by storing latent energy at the melting temperature of a lipid-based engineered phase change material. Endpoint classification of the reaction is achieved without opening the reaction tube by observing the fluorescence of sequence-specific FRET-based assimilating probes with a simple handheld fluorometer. At or above 22°C ambient temperature the non-instrumented devices could maintain reactions above a threshold temperature of 61°C for over 90 min-significantly longer than the 60 min reaction time. Using the simple format, detection limits were less than 20 genome copies for reactions run at ambient temperatures ranging from 8 to 36°C. When used with a pre-enrichment step and non-instrumented DNA extraction device, trace contaminations of Salmonella in milk close to 1 CFU/mL could be reliably detected. These findings illustrate that the non- instrumented amplification approach is a simple, viable, low-cost alternative for field-based food and agricultural diagnostics or clinical applications in developing countries.</p>1 aKubota, Ryo1 aLaBarre, Paul1 aWeigl, Bernhard, H1 aLi, Yong1 aHaydock, Paul1 aJenkins, Daniel, M  uhttp://microfluidicsciences.com/drupal/?q=node/204