Simple LAMP PCR design for low resource setting and minimal environmental footprint
Inventors
Beitle, Robert • Hestekin, Christa N. • Elmasheiti, Ahmed • Cribbs, Kimberly • Rienisch, Michael • Jones, Bryce Cameron • Swearingen, Allysa • Hart, Brandon • Clark, Kevin • Thompson, Vicki
Assignees
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Publication Number
US-11052330-B2
Publication Date
2021-07-06
Expiration Date
Abstract
The present invention provides a device and method for testing a material for the presence of DNA. The system includes a centrifuge, a microchip performing cell lysis and an enclosure that contains an isothermal ballast material and chromogenic agent that melts at a specific temperature and displays a color change, respectively.
Core Innovation
The invention provides a low-resource DNA testing method in which a sample containing cells is placed in an enclosure, the cells are concentrated and lysed, and DNA is extracted to form a second analyte. The second analyte is amplified by applying heat at a predetermined temperature, and DNA is detected after amplification in the enclosure.
Temperature maintenance during heating is achieved using a phase change material that melts at the predetermined temperature to maintain the predetermined temperature. In a further implementation, the enclosure is lined with a phase change material that includes a thermochromic pigment that changes color at a temperature equal to or greater than the predetermined temperature.
The disclosure also describes DNA extraction by adsorption onto silica using chaotropic salt solutions, followed by DNA detection using dyeing. A specific visualization technique uses an SYBR green technique after DNA extraction and amplification, and the testing method for the presence of DNA is performed without the use of electric current by applying chemical heating to the enclosure.
Claims Coverage
Two independent claims are identified: one directed to amplifying and detecting DNA using heat maintained by a phase change material, and one directed to testing for DNA presence without electric current using a phase change material that includes a thermochromic pigment. Across the independent claims, the inventive core includes concentrating cells, lysing, extracting DNA, heating-amplifying DNA in an enclosure, and isothermal temperature maintenance using a phase change material, with downstream DNA detection.
Enclosure temperature maintained by phase change material during DNA amplification
A method for testing a material that includes applying heat at a predetermined temperature to an enclosure to amplify DNA in a second analyte, using a phase change material that melts at a predetermined temperature to maintain the predetermined temperature during said heating; and detecting said DNA.
Heating with thermochromic pigment in phase change material for visual indication without electric current
A method for testing a material for the presence of DNA without the use of an electric current by obtaining a sample of cells, concentrating said cells, lysing said cells, extracting DNA to form a second analyte, said second analyte being heated at a predetermined temperature in an enclosure lined with a phase change material that melts at a predetermined temperature to maintain said predetermined temperature during heating and includes a thermochromic pigment that changes color at a temperature equal to or greater than said predetermined temperature; and detecting said DNA.
The independent claims collectively cover isothermal DNA amplification within an enclosure where temperature is maintained by a phase change material that melts at the predetermined temperature, with one independent claim including thermochromic pigment coloration and explicitly requiring no electric current.
Stated Advantages
Maintains a predetermined temperature during DNA amplification by using a phase change material that melts at the predetermined temperature.
Provides a visual color change indicating reaching a temperature equal to or greater than the predetermined temperature via a thermochromic pigment.
Enables testing for the presence of DNA without the use of an electric current.
Documented Applications
Testing a material by concentrating cells, lysing, extracting DNA, amplifying DNA at a predetermined temperature maintained using a melting phase change material, and detecting the DNA.
Testing a material for the presence of DNA without the use of an electric current by heating in an enclosure lined with a phase change material that includes a thermochromic pigment for color change at the predetermined temperature or higher.
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