Heating mechanism for DNA amplification, extraction or sterilization using photo-thermal nanoparticles
A heating mechanism for use in DNA applications such as DNA amplification, extraction and sterilization is provided. Nanoparticles having photo-thermal properties are put in contact with a reaction mixture and irradiated with an activation light beam to activate these photo-thermal properties, thereby releasing heat. Nanoparticles of several types may be used. Use of the same nanoparticles or of different one to monitor the reaction using a different light beam is also presented.
1. A method of amplifying a nucleic acid molecule with a polymerase chain reaction (PCR) or a loop-mediated isothermal amplification (LAMP), through bulk heating a biological enzymatic reaction mixture in solution containing
a nucleic acid template comprising a reverse transcribed nucleic acid,
a polymerase enzyme, and
chemically modified nanoparticles comprising nanorods of metal, metallic coated organic nanotubes, or a combination thereof, having photo-thermal properties, to promote said LAMP, comprising:
a) irradiating said chemically modified nanoparticles with an activation light beam from a continuous wave laser to provide excitation for a period of time to activate said photo-thermal properties of said chemically modified nanoparticles, such that said chemically modified nanoparticles release heat sufficient to provide said heating to the whole reaction mixture in solution and promote said PCR or said LAMP.
2. The method according claim 1 , wherein the nanoparticles are selected from the group consisting of carbon nanotubes coated with a metal and multiwalled carbon nanotubes coated with or decorated with a metal.
3. The method according to claim 2 , wherein the metal is selected from the group consisting of Au, Ag, Pd, Pt, Fe, Cu, Al, and Zn.
4. The method according to claim 1 , wherein said photo-thermal properties comprise a localized plasmon resonance at a surface of the chemically modified nanoparticles, and the activation light beam has a wavelength corresponding to said localized plasmon resonance.
5. The method according to claim 1 , wherein the chemically modified nanoparticles are chemically modified by a chemical compound that prevents the inhibition of an active site of said polymerase enzyme.
6. The method according to claim 5 , wherein the chemical compound that prevents the inhibition of the active site of said polymerase enzyme is polyethylene glycol.
7. The method according to claim 1 , wherein the step of irradiating comprises adjusting a power of said activation light beam to regulate temperature of said biological enzymatic reaction mixture in solution through controlled heat release from said chemically modified nanoparticles.
8. The method according to claim 1 , further comprising a step of monitoring said bulk heating, amplicon production, or both.
9. The method according to claim 8 , wherein the step of monitoring said amplicon production comprises probing said chemically modified nanoparticles with a probing light beam, having a wavelength different than a wavelength of the activation light beam and coordinated with an absorption feature of said chemically modified nanoparticles spectrally separate from the photo-thermal properties used to release heat, to measure a change of an optical property of said chemically modified nanoparticles and correlate said change of the optical property with a change in said amplicon production.
10. The method according to claim 9 , wherein said chemically modified nanoparticles have an elongated geometry, the wavelength of the activation light beam is coordinated with a longitudinal resonance of the nanoparticles and the wavelength of the probing light beam is coordinated with a transversal resonance of the nanoparticles.
11. The method of claim 1 , comprising cooling of the reaction mixture after the heating thereof.
12. The method of claim 1 , further comprising the step of reverse transcription of a nucleic acid to provide said nucleic acid template comprising a reverse transcribed nucleic acid prior to amplifying said nucleic acid molecule with PCR or LAMP.
13. The method of claim 12 , further comprising the step of extracting an RNA from a cell, a virus or bacteria, prior to said step of reverse transcription.
14. The method of claim 13 , wherein said step of extracting RNA from a cell, a virus or bacteria comprises irradiating said chemically modified nanoparticles with an activation light beam from a continuous wave laser to provide excitation for a period of time to activate said photo-thermal properties of said chemically modified nanoparticles, such that said chemically modified nanoparticles release heat sufficient for extraction of said RNA from said cell, virus or bacteria.
15. The method of claim 14 , further comprising the step of reverse transcription of a nucleic acid to provide said nucleic acid template comprising a reverse transcribed nucleic acid prior to amplifying said nucleic acid molecule with PCR or LAMP.
16. The method of claim 15 , further comprising the step of extracting an RNA from a cell or a virus prior to said step of reverse transcription.
17. The method of claim 16 , wherein said step of extracting said RNA from a cell or a virus or bacteria comprises irradiating said chemically modified nanoparticles with an activation light beam from a continuous wave laser to provide excitation for a period of time to activate said photo-thermal properties of said chemically modified nanoparticles, such that said chemically modified nanoparticles release heat sufficient for extraction of said RNA from said cell, virus or bacteria.
18. A method of amplifying a nucleic acid molecule with a polymerase chain reaction (PCR) or loop-mediated isothermal amplification (LAMP) through bulk heating a biological enzymatic reaction mixture in solution containing
a nucleic acid template comprising a reverse transcribed nucleic acid,
a polymerase enzyme,
first chemically modified nanoparticles comprising nanorods of metal, metallic coated organic nanotubes, or a combination thereof, having photo-thermal properties, to release heat and promote said PCR or said LAMP, and
a second set of nanoparticles having an absorption feature spectrally separate from said photo-thermal properties of said first chemically modified nanoparticles,
comprising the steps of:
a) irradiating said first chemically modified nanoparticles with an activation light beam from a continuous wave laser to provide excitation for a period of time to activate said photo-thermal properties of said first chemically modified nanoparticles, such that said first chemically modified nanoparticles release heat sufficient to provide said heating to the whole reaction mixture in solution and promote said PCR or said LAMP, and
b) monitoring said bulk heating, amplicon production, or both, by probing said second set of nanoparticles with a probing light beam having a wavelength different than a wavelength of the activation light beam and coordinated with said absorption feature.
19. The method of claim 18 , wherein said second set of nanoparticles is second chemically modified nanoparticles comprising nanorods of metal, metallic coated organic nanotubes, or a combination thereof, having photo-thermal properties, and having an absorption feature spectrally separate from said photo-thermal properties of said first chemically modified nanoparticles.
20. The method of claim 8 , wherein said nucleic acid template is from a cell, a virus or bacteria.