Detection of negatively charged polymers using water-soluble, cationic, polythiophene derivatives
View Patent ↗Novel methods allowing for the simple optical and electrochemical detection of double-stranded oligonucleotides are disclosed. The methods are rapid, selective and versatile. Advantageously, they do not require any chemical reaction on the probes or on the analytes since they are based on different electrostatic interactions between cationic poly (3-alkoxy-4-methylthiophene) derivatives and single-stranded or double-stranded (hibridized) oligonucleotides.
1. A method for detecting the presence of a negatively charged polymer, comprising:
a) contacting a target complementary to said negatively charged polymer with a cationic polymer to form a duplex, wherein the cationic polymer comprises the following formula:
wherein:
m is an integer ranging from 2 to 3;
n is an integer ranging from 3 to 100;
R* is a quartenary ammonium;
Y is an oxygen atom or a methylene; and
R 1 is a methyl group or a hydrogen atom;
b) contacting said duplex with said negatively charged polymer; and
c) detecting a change in electronic charge, fluorescence or color as an indication of the presence of said negatively charged polymer.
2. The method as defined in claim 1 , wherein said negatively charged polymer is selected from the group consisting of: an acidic protein, a glycosaminoglycan, a hyaluronan, heparin, a chromatographic substrate, a culture substrate and a nucleic acid.
3. The method as defined in claim 2 , wherein said negatively charged polymer is a nucleic acid.
4. The method as defined in claim 3 , wherein said nucleic acid is DNA.
5. The method as defined in claim 3 , wherein said nucleic acid is RNA.
6. The method as defined in claim 3 , wherein said complementary target is a nucleic acid complementary to said negatively charged polymer.
7. The method as defined in claim 6 , wherein said complementary target is a nucleic acid probe and said negatively charged polymer is a denatured PCR amplicon.
8. The method of claim 1 , wherein said cationic polymer is ionically linked to said target complementary to said negatively charged polymer.
9. The method of claim 1 , wherein said target complementary to said negatively charged polymer is linked to a solid support.
10. The method of claim 9 , further comprising reacting said positively charged polymer with said target complementary to said negatively charged polymer, resulting in capture of said target complementary to said negatively charged polymer by said solid support forming a complex, and reacting said negatively charged polymer with said complex.
11. The method of claim 9 , wherein said solid support is selected from the group consisting of an electrode, an optical fiber, a glass slide and glass beads.
12. The method of claim 9 , further comprising purifying said negatively charged polymers.
13. The method of claim 12 , wherein said negatively charged polymers are nucleic acids.
14. The method of claim 9 , further comprising reacting said negatively charged polymer with said target complementary to said negatively charged polymer to form a complex between said negatively charged polymer and target complementary to said negatively charged polymer, wherein said positively charged polymer is reacted with said complex.
15. The method of claim 1 , wherein:
m is 3;
R* is + Net 3 ;
Y is an oxygen atom; and
R 1 is a methyl group.
16. The method of claim 1 , wherein:
m is 2;
R* is
Y is an oxygen atom; and
R 1 is a methyl group.
17. The method of claim 1 , wherein
m is 2;
R* is
Y is an oxygen atom; and
R 1 is a methyl group.
18. The method of claim 1 , wherein
m is 2;
R* is
Y is a methylene group; and
R 1 is a hydrogen atom.
19. The method of claim 1 , wherein the polymer formula is selected from the group consisting of: