02745nas a2200373 4500008004100000022001400041245009900055210006900154260000900223300001100232490000600243520168200249653002301931653001601954653001501970653002101985653001202006653001902018653001002037653001102047653004202058653002602100653001502126653001602141100002102157700002202178700001702200700002102217700001702238700002002255700001802275700001802293856006002311 2012 eng d a1932-620300aIsothermal amplification using a chemical heating device for point-of-care detection of HIV-1.0 aIsothermal amplification using a chemical heating device for poi c2012 ae314320 v73 a

BACKGROUND: To date, the use of traditional nucleic acid amplification tests (NAAT) for detection of HIV-1 DNA or RNA has been restricted to laboratory settings due to time, equipment, and technical expertise requirements. The availability of a rapid NAAT with applicability for resource-limited or point-of-care (POC) settings would fill a great need in HIV diagnostics, allowing for timely diagnosis or confirmation of infection status, as well as facilitating the diagnosis of acute infection, screening and evaluation of infants born to HIV-infected mothers. Isothermal amplification methods, such as reverse-transcription, loop-mediated isothermal amplification (RT-LAMP), exhibit characteristics that are ideal for POC settings, since they are typically quicker, easier to perform, and allow for integration into low-tech, portable heating devices.

METHODOLOGY/SIGNIFICANT FINDINGS: In this study, we evaluated the HIV-1 RT-LAMP assay using portable, non-instrumented nucleic acid amplification (NINA) heating devices that generate heat from the exothermic reaction of calcium oxide and water. The NINA heating devices exhibited stable temperatures throughout the amplification reaction and consistent amplification results between three separate devices and a thermalcycler. The performance of the NINA heaters was validated using whole blood specimens from HIV-1 infected patients.

CONCLUSION: The RT-LAMP isothermal amplification method used in conjunction with a chemical heating device provides a portable, rapid and robust NAAT platform that has the potential to facilitate HIV-1 testing in resource-limited settings and POC.

10aAIDS Serodiagnosis10aDNA Primers10aDNA, Viral10aEquipment Design10aHeating10aHIV Infections10aHIV-110aHumans10aNucleic Acid Amplification Techniques10aPoint-of-Care Systems10aRNA, Viral10aTemperature1 aCurtis, Kelly, A1 aRudolph, Donna, L1 aNejad, Irene1 aSingleton, Jered1 aBeddoe, Andy1 aWeigl, Bernhard1 aLaBarre, Paul1 aOwen, Michele uhttps://www.microfluidicsciences.com/drupal/?q=node/20701963nas a2200217 4500008004100000022001400041245011800055210006900173260000900242300001000251490000600261520127900267100001601546700001801562700002101580700001701601700002301618700002101641700002301662856006001685 2011 ENG d a2162-643X00aNon-Instrumented Nucleic Acid Amplification (NINA) for Rapid Detection of Ralstonia solanacearum Race 3 Biovar 2.0 aNonInstrumented Nucleic Acid Amplification NINA for Rapid Detect c2011 a69-800 v43 a

We report on the use of a non-instrumented device for the implementation of a loop-mediated amplification (LAMP) based assay for the select-agent bacterial-wilt pathogen Ralstonia solanacearum race 3 biovar 2. Heat energy is generated within the device by the exothermic hydration of calcium oxide, and the reaction temperature is regulated by storing latent energy at the melting temperature of a renewable lipid-based engineered phase-change material. Endpoint detection of the LAMP reaction is achieved without opening the reaction tube by observing the fluorescence of an innovative FRET-based hybridization probe with a simple custom fluorometer. Non-instrumented devices could maintain reactions near the design temperature of 63°C for at least an hour. Using this approach DNA extracted from the pathogen could be detected at fewer than ten copies within a 25 μL reaction mix, illustrating the potential of these technologies for simple, powerful agricultural diagnostics in the field. Furthermore, the assay was just as reliable when implemented in a tropical environment at 31°C as it was when implemented in an air-conditioned lab maintained at 22°C, illustrating the potential value of the technology for field conditions in the tropics and subtropics.

1 aKubota, Ryo1 aLaBarre, Paul1 aSingleton, Jered1 aBeddoe, Andy1 aWeigl, Bernhard, H1 aAlvarez, Anne, M1 aJenkins, Daniel, M uhttps://www.microfluidicsciences.com/drupal/?q=node/214