Apparatuses and methods for detecting nucleic acids
View Patent ↗The present disclosure provides a low cost, high sensitivity, high specificity and rapid diagnostic apparatus and method to detect nucleic acids in a sample at room temperature. As low as tens of copies of nucleic acids can be detected without any additional equipment.
1. An apparatus comprising:
(a) a surface for attaching a DNA capture probe;
(b) the capture DNA probe;
(c) a target nucleic acid, wherein a first portion of the target nucleic acid sequence is partially complementary to the sequence of the capture DNA probe;
(d) a nanoparticle for detecting nucleic acids comprising (i) a nanoparticle; (ii) a radical initiator attached to the nanoparticle; and (iii) a detector DNA attached to the nanoparticle, wherein the detector DNA has a sequence that is partially complementary to a second portion of sequence of a target nucleic acid; and
(e) a monomer, wherein the monomer is (ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl) bis(2-methylacrylate).
2. A method for preparing the apparatus of claim 1 , the method comprising:
(a) attaching a capture DNA probe to a surface, wherein the capture DNA probe comprises a functional group capable of attaching to the surface and comprises a sequence that is partially complementary to a first portion of a sequence of target nucleic acid;
(b) contacting the target nucleic acid with the capture DNA probe attached to the surface from step (a), wherein the first portion of the target nucleic acid partially hybridizes to the capture DNA probe;
(c) contacting a nanoparticle for detecting nucleic acids with the product of step (b), wherein the nanoparticle comprises (i) a nanoparticle; (ii) a radical initiator attached to the nanoparticle; and (iii) a detector DNA probe attached to the nanoparticle, the detector DNA probe comprising a sequence that is partially complementary to a second portion of the sequence of target nucleic acid, and wherein the detector DNA probe of the nanoparticle partially hybridizes to the second portion of the target nucleic acid; and
(d) contacting the product of step (c) with a monomer, wherein the monomer is (ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl) bis(2-methylacrylate).
3. A method for detecting nucleic acid in a sample, the method comprising:
(a) contacting a monomer with the apparatus of the claim 1 ;
(b) exposing the product of step (a) to light; and
(c) detecting the presence of a polymer, wherein the presence of polymer indicates nucleic acid in the sample;
wherein the nucleic acid is detected by a visual signal of polymerization of the monomer.
4. An apparatus comprising:
(a) a surface for attaching a DNA capture probe;
(b) the capture DNA probe;
(c) a target nucleic acid, wherein a first portion of the target nucleic acid sequence is partially complementary to the sequence of the capture DNA probe;
(d) a nanoparticle for detecting nucleic acids comprising (i) a nanoparticle; (ii) a radical initiator attached to the nanoparticle; and (iii) a detector DNA attached to the nanoparticle, wherein the detector DNA has a sequence that is partially complementary to a second portion of sequence of a target nucleic acid; and
(e) a polymer, wherein the polymer is a compound of formula 10:
wherein
Y is independently selected from O, S, NH or NR 1 ;
R is independently selected from H, (C 2 -C 6 ) alkyne, R 2 SO 3 , (C 1 -C 20 ) polyhydroxy alkyl, (C 1 - 20 ) polyamino alkyl or carbohydrate; and
R 1 and R 2 are independently H or (C 1 -C 6 ) alkyl; and
n is an integer selected from 2 to 2000.
5. An apparatus comprising:
(a) a surface for attaching a DNA capture probe;
(b) the capture DNA probe;
(c) a target nucleic acid, wherein a portion of the target nucleic acid sequence is partially complementary to the sequence of the capture DNA probe;
(d) a compound of formula (I), wherein the compound of formula (I):
A-B-C (I),
wherein:
A is an initiator;
B is a linker; and
C is a detector DNA probe, the DNA detector probe comprising a sequence that is capable of partially hybridizing to a target nucleic acid; and
(e) a monomer, wherein the monomer is (ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl) bis(2-methylacrylate).
6. A method for preparing the apparatus of claim 5 , the method comprising:
(a) attaching a capture DNA probe to a surface, wherein the capture DNA probe comprises a functional group capable of attaching to the surface and comprises a sequence that is partially complementary to a first portion of a sequence of target nucleic acid;
(b) contacting the target nucleic acid with the capture DNA probe attached to the surface from step (a), wherein the first portion of the target nucleic acid partially hybridizes to the capture DNA probe;
(c) contacting the compound of formula (I) with the product of step (b), wherein the detector DNA probe partially hybridizes to the second portion of the target nucleic acid and wherein the compound of formula (I) is:
A-B-C (I),
wherein:
A is an initiator;
B is a linker; and
C is a detector DNA probe, the DNA detector probe comprising a sequence that is capable of partially hybridizing to a target nucleic acid; and
(d) contacting the product of step (c) with a monomer, wherein the monomer is (ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl) bis(2-methylacrylate).
7. An apparatus comprising:
(a) a surface for attaching a DNA capture probe;
(b) the capture DNA probe;
(c) a target nucleic acid, wherein a portion of the target nucleic acid sequence is partially complementary to the sequence of the capture DNA probe;
(d) a compound of formula (I), wherein the compound of formula (I):
A-B-C (I),
wherein:
A is an initiator;
B is a linker; and
C is a detector DNA probe, the DNA detector probe comprising a sequence that is capable of partially hybridizing to a target nucleic acid; and
(e) a polymer, wherein the polymer is compound of formula 10:
wherein
Y is independently selected from O, S, NH or NR 1 ;
R is independently selected from H, (C 2 -C 6 ) alkyne, R 2 SO 3 , (C 1 -C 20 ) polyhydroxy alkyl, (C 1 -C 20 ) polyamino alkyl or carbohydrate; and
R 1 and R 2 are independently H or (C 1 -C 6 ) alkyl; and
n is an integer selected from 2 to 2000.
8. An apparatus comprising:
(a) a surface for attaching a DNA capture probe;
(b) the capture DNA probe;
(c) a target nucleic acid, wherein a portion of the target nucleic acid sequence is partially complementary to the sequence of the capture DNA probe;
(d) a compound of formula (I), wherein the compound of formula (I):
A-B-C (I),
wherein:
A is an initiator;
B is a linker; and
C is a detector DNA probe, the DNA detector probe comprising a sequence that is capable of partially hybridizing to a target nucleic acid; and
(e) a polymer, wherein the polymer comprises (ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl) bis(2-methylacrylate) monomer.
9. A method for detecting nucleic acid in a sample, the method comprising:
(a) contacting the monomer (ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl) bis(2-methylacrylate) with an apparatus, the apparatus comprising:
(i) a surface for attaching a DNA capture probe;
(ii) the capture DNA probe;
(iii) a target nucleic acid, wherein a portion of the target nucleic acid sequence is partially complementary to the sequence of the capture DNA probe; and
(iv) a compound of formula (I):
A-B-C (I),
wherein:
A is an initiator;
B is a linker; and
C is a detector DNA probe, the DNA detector probe comprising a sequence that is capable of partially hybridizing to a target nucleic acid;
(b) exposing the product of step (a) to light; and
(c) detecting the presence of a polymer, wherein the presence of polymer indicates nucleic acid in the sample;
wherein the nucleic acid is detected by a visual signal of polymerization of the monomer.