Microfluidic platforms for multi-target detection
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12Q-001/68
C12P-019/34
C12M-001/36
C12M-001/34
G01N-015/06
G01N-027/00
B01J-019/08
B01D-057/02
B03C-005/02
C02F-001/48
C07H-021/02
C07H-021/04
출원번호
US-0246987
(2008-10-07)
등록번호
US-8771938
(2014-07-08)
발명자
/ 주소
Chang, Hsueh-Chia
Gordon, Jason
Senpati, Satyajyoti
Gagnon, Zachary
Basuray, Sagnik
출원인 / 주소
University of Notre Dame du Lac
대리인 / 주소
Greenberg Traurig, LLP
인용정보
피인용 횟수 :
1인용 특허 :
5
초록▼
Disclosed are example methods and devices for detecting one or more targets. An example method includes placing a sample including a first target with in a microfluidic device and hybridizing a plurality of copies of the first target with a plurality of nanostructures. The example method includes ap
Disclosed are example methods and devices for detecting one or more targets. An example method includes placing a sample including a first target with in a microfluidic device and hybridizing a plurality of copies of the first target with a plurality of nanostructures. The example method includes applying an electric current to the plurality of nanostructures and using an electric field created by the electric current to move the plurality of nanostructures. In addition, the plurality of nanostructures are sorted and evaluated to determine at least one of a presence, an absence, or a quantity of the first target.
대표청구항▼
1. A method of detecting a plurality of target nucleic acids, the method comprising: continuously flowing a sample solution comprising the target nucleic acids through a microfluidic device having at least a first surface with a first electrode and a second surface displaced from the first surface a
1. A method of detecting a plurality of target nucleic acids, the method comprising: continuously flowing a sample solution comprising the target nucleic acids through a microfluidic device having at least a first surface with a first electrode and a second surface displaced from the first surface at a distance and having a second electrode opposing the first electrode;provided a first and second functionalized nanostructure in solution to the microfluidic device, wherein the first nanostructure is formed by functionalizing a first probe complementary to a first target nucleic acid and the second nanostructure is formed by functionalizing a second probe complementary to a second target nucleic acid;applying an electric current to the electrodes;using the electric field created by the electric current applied to the electrodes to move the nanostructures;trapping the nanostructures within the electric field;mixing the sample solution and the solution providing the nanostructure;hybridizing the target nucleic acids with the nanostructures in the presence of the electric field;hydrodynamically shearing at least one of a non-target or a weakly hybridized target nucleic acid from the nanostructures, wherein the application of the electric current to the nanostructures causes the nanostructure hybridized with the first target to move in a first direction and causes the nanostructure hybridized with the second target to move in a second direction, and wherein the first target and the second target are sorted based on the first direction or the second direction; andevaluating the trapped nanostructures by measuring an electrical impedance between the electrodes to determine a presence, an absence, or a quantity of the first or second target nucleic acids. 2. A method as defined in claim 1, wherein the electric current induces dielectrophoresis. 3. A method as defined in claim 1, wherein a presence of an increased pressure does not impede the method. 4. A method as defined in claim 1, wherein a copy of at least one of the plurality of target nucleic acids is produced via a polymerase chain reaction. 5. A method as defined in claim 1, wherein the electric field has at least a first frequency and a second frequency, and wherein the nanostructures move in a first direction at the first frequency and in a second direction at the second frequency. 6. A method as defined in claim 1, wherein the plurality of nanostructures form a pattern dependent on a frequency of the electric field. 7. A method as defined in claim 1, wherein the electric current creates a non-uniform electric field across the microfluidic device. 8. A method as defined in claim 1, wherein the nanostructures are one or more of carbon nanotubes, nanobeads, nanowires, nanocolloides, nanoparticles, nanorods, quantum dots, nanocrystals, liposomes, silica beads, latex beads, gold colloids or other structures with dimensions less than one micron. 9. A method as defined in claim 1, wherein the one or more of the first or second probes includes one or more of a oligomer, a fluorophore, a carboxyl group, or a streptavidin. 10. A method as defined in claim 1 further comprising pretreating the sample. 11. A method as defined in claim 10, wherein pretreating includes at least one of filtering or removal of inhibitors. 12. A method of detecting a target nucleic acid, the method comprising: obtaining a sample including the target nucleic acid;functionalizing a first molecular probe complementary to a first target nucleic acid to a first nanostructure and functionalizing a second molecular probe complementary to a second target nucleic acid to a second nanostructure;coupling the functionalized nanostructures to a chamber having a first surface with a first electrode and a second surface displaced from the first surface at a distance and having a second electrode opposing the first electrode;flowing the target nucleic acids through the chamber to hybridize the first target nucleic acid to the first functionalized nanostructure and to hybridize the second target nucleic acid to the second functionalized nanostructure;causing the nanostructure hybridized with the first target to move in a first direction and causing the nanostructure hybridized with the second target to move in a second direction, wherein the first target and the second target are sorted based on the first direction or the second direction; and detecting at least one of a presence, an absence, or a quantity of the first or second target nucleic acids by measuring an electrical impedance between the electrodes. 13. A method as defined in claim 12, wherein at least one of the first or second target nucleic acids is replicated through a polymerase chain reaction and flowing the amplified mixture through the chamber. 14. A method as defined in claim 13, wherein the polymerase chain reaction uses two differently labeled primers. 15. A method as defined in claim 14, wherein one of the primers is biotinylated and the other is fluorescently labeled. 16. A method as defined in claim 12 further comprising applying an alternating current electric field to the chamber wherein the presence of the electric field improves hybridization rate and yield. 17. A method of detecting target nucleic acids comprising: placing a solution having a plurality of functionalized nanostructures within a channel having a first surface with a first electrode and a second surface displaced from the first surface at a distance and having a second electrode opposing the first electrode, wherein a first one of the plurality of functionalized nanostructures includes a oligonucleotide probe complementary to a first target nucleic acid and wherein a second one of the plurality of functionalized nanostructures includes a oligonucleotide probe complementary to a second target nucleic acid;applying an alternating current field to the first and second electrodes to focus and trap the nanostructures;flowing a sample including the target nucleic acids within the channel and through the trapped functionalized nanostructures;hybridizing the target nucleic acids with at least one of the nanostructures;hydrodynamically shearing at least one of a non-target or a weakly hybridized target nucleic acid from the nanostructures, wherein the application of the electric current to the nanostructures causes the nanostructure hybridized with the first target to move in a first direction and causes the nanostructure hybridized with the second target to move in a second direction, and wherein the first target and the second target are sorted based on the first direction or the second direction; andevaluating the trapped nanostructures by measuring an impedance signal during the sample flow to determine a presence, an absence, or a quantity of the target nucleic acids. 18. A method as defined in claim 17, wherein the applied electric field is an alternating current field with a period shorter than the Faradaic reaction time corresponding to the respective voltage. 19. A method as defined in claim 18, wherein the alternating current applied to the first and second electrodes creating the electric field is also used to provide the measurement of electrical impedance between the electrodes. 20. A method as defined in claim 17, wherein the nanostructures are carbon nanotubes. 21. A method as defined in claim 20, wherein the carbon nanotubes trapped at the electrodes focus the electric field thereby concentrating targets between the first and second electrodes. 22. A method as defined in claim 21, wherein the carbon nanotubes enhance dielectropheretic forces within the channel. 23. A method as defined in claim 17, wherein hybridization yield and rate are enhanced in the presences of an electric field. 24. A method as defined in claim 17, wherein the nanostructures are trapped by force induced by dielectrophoresis. 25. A method as defined in claim 17, wherein the nanostructures are evaluated based on a change in impedance signal before and after hybridization with the target nucleic acids with the shift in impedance relevant to the number of hybridized targets.
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이 특허에 인용된 특허 (5)
Wu Annie L. (Penfield NY), Detection of amplified nucleic acid using secondary capture oligonucleotides and test kit.
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