@article {207, title = {Isothermal amplification using a chemical heating device for point-of-care detection of HIV-1.}, journal = {PLoS One}, volume = {7}, year = {2012}, month = {2012}, pages = {e31432}, abstract = {

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.

}, keywords = {AIDS Serodiagnosis, DNA Primers, DNA, Viral, Equipment Design, Heating, HIV Infections, HIV-1, Humans, Nucleic Acid Amplification Techniques, Point-of-Care Systems, RNA, Viral, Temperature}, issn = {1932-6203}, doi = {10.1371/journal.pone.0031432}, author = {Curtis, Kelly A and Rudolph, Donna L and Nejad, Irene and Singleton, Jered and Beddoe, Andy and Weigl, Bernhard and LaBarre, Paul and Owen, S Michele} } @article {214, title = {Non-Instrumented Nucleic Acid Amplification (NINA) for Rapid Detection of Ralstonia solanacearum Race 3 Biovar 2.}, journal = {Biol Eng Trans}, volume = {4}, year = {2011}, month = {2011}, pages = {69-80}, abstract = {

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{\textdegree}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{\textdegree}C as it was when implemented in an air-conditioned lab maintained at 22{\textdegree}C, illustrating the potential value of the technology for field conditions in the tropics and subtropics.

}, issn = {2162-643X}, doi = {10.13031/2013.38508}, author = {Kubota, Ryo and LaBarre, Paul and Singleton, Jered and Beddoe, Andy and Weigl, Bernhard H and Alvarez, Anne M and Jenkins, Daniel M} }