Probe for detecting and treating virus

Patent metadata

Field	Value
Publication number	US-11535901-B2
Application number	US-201816142073-A
Country	US
Kind code	B2
Filing date	Sep 26, 2018
Priority date	Sep 26, 2017
Publication date	Dec 27, 2022
Grant date	Dec 27, 2022

Abstract

Provided herein are a probe capable of simultaneously detecting a virus and treating virus-infected cells, a composition for detecting a virus, which comprises the probe, a composition for treating a virus, which comprises the probe, and a method of detecting a virus or treating a viral infection by using the same. According to the present disclosure, it is possible to simultaneously perform diagnosis by virus detection and treatment of virus-infected cells, and in particular, diagnosis and treatment may be simultaneously performed on various types of viruses by varying the type of molecular beacon, and thus may be usefully applied to virus diagnosis and treatment fields, which require rapid diagnosis and treatment, and the spread of viral infections may be effectively prevented. In addition, the probe of the present disclosure has excellent stability and excellent detection sensitivity, and thus enables the detection of even a very low amount of a target at the pmole level.

First claim

What is claimed is:

1. A probe for detecting and/or treating a virus, the probe comprising:
a metal particle; and
one or more molecular beacons, each molecular beacon comprising an anti-target nucleic acid molecule and an oligo nucleic acid molecule,
wherein the anti-target nucleic acid molecule is bound to the surface of the metal particle and comprises a non-target site and a target-specific binding site;
wherein the oligo nucleic acid molecule comprises a non-target site, an anti-target nucleic acid molecule-complementary binding site and a fluorescent material;
wherein the GC content in the non-target site of the anti-target nucleic acid molecule is 50% or more and the GC content in the non-target site of the oligo nucleic acid molecule is 50% or more;
wherein the non-target site of the oligo nucleic acid molecule complementarily binds to the non-target site of the anti-target nucleic acid molecule;
wherein the anti-target nucleic acid molecule and the oligo nucleic acid molecule are two separate nucleic acid strands,
wherein the probe has a temperature (Tm 0 ) greater than or equal to 36° C. (Tm 0 ≥36° C.), the temperature (Tm 0 ) being a temperature at which for a population of identical molecular beacons, the anti-target nucleic acid molecule and the oligo nucleic acid molecule of 50% of the total molecular beacons are separated from each other;
wherein the probe has a temperature (Tm 1 ) that is lower than Tm 0 (Tm 0 >Tm 1 ), the temperature (Tm 1 ) being a temperature at which for a population of identical molecular beacons, when the target-specific binding site of the anti-target nucleic acid molecule is bound to a target, 50% of the total target-specific binding site is separated from the target; and
wherein the probe has a temperature (Tm 2 ) that is lower than Tm 0 and Tm 1 , the temperature (Tm 2 ) being a temperature at which for a population of identical molecular beacons, when the target-specific binding site of the anti-target nucleic acid molecule is bound to a target, 50% of a non-target site of the anti-target nucleic acid molecule is separated from a non-target site of the oligo nucleic acid molecule.

2. The probe of claim 1 , wherein the probe comprises two or more molecular beacons of two or more types,
wherein the target of each of the anti-target nucleic acid molecules in the two or more molecular beacons are different from each other, and
the fluorescent material of each of the oligo nucleic acid molecules in the two or more molecular beacons has a different emission wavelength.

3. The probe of claim 1 , wherein the target-specific binding site of the anti-target nucleic acid molecule complementarily binds to a nucleic acid sequence encoding a stem region in a hemagglutinin (HA) protein of an influenza virus.

4. The probe of claim 3 , wherein the virus comprises one or more selected from the group consisting of H1N1, H2N2, H3N2, H5N1, H7N7, and H9N2.

5. The probe of claim 1 , wherein the anti-target nucleic acid molecule comprises a non-target site and a target-specific binding site having a sequence of 2 to 24 nucleotides, the sequence complementarily binds to any one nucleic acid sequence selected from the group consisting of SEQ ID NOS: 1 to 3.

6. The probe of claim 1 , wherein the metal is one or more selected from the group consisting of gold (Au), silver (Ag), platinum (Pt), and palladium (Pd).

7. The probe of claim 1 , wherein the fluorescent material is one or more selected from the group consisting of rhodamine or derivatives thereof, fluorescein or derivatives thereof, coumarin or derivatives thereof, acridine and derivatives thereof, pyrene and derivatives thereof, erythrosine and derivatives thereof, eosin or derivatives thereof, cyanine or derivatives thereof, and 4-acetamido-4′-isothiocyanatostilbene-2,2′-disulfonic acid.

8. The probe of claim 1 , wherein the virus is an influenza virus.

9. A composition for detecting a virus comprising the probe according to claim 1 , wherein the composition comprises two or more different types of probes that detect different targets.

10. A method of treating a viral infection, the method comprising:
administering to a virus-infected subject the probe according to claim 1 .

11. The method of claim 10 , wherein the probe comprises two or more molecular beacons.

12. A method of detecting a virus, the method comprising:
contacting a sample obtained from a subject with the probe according to claim 1 ; and
measuring a change in fluorescence of the sample contacted with the probe.

Assignees

• Yonsei Univ Industry Foundation Yonsei Uif
• Univ Korea Res & Business Foundation Sejong Campus

Inventors

• Haam Seung Joo
• Kim Hyun Ouk
• Kang Byung Hoon
• Han Seung Min
• Ki Ji Sun
• Song Dae Sub
• Na Woon Sung
• Yeom Min Joo

Classifications

• C12Q1/702 Primary

  for retroviruses · CPC title

• C12Q1/701 Primary

  Specific hybridization probes · CPC title

• C12Q2563/107

  fluorescence · CPC title

• C12Q1/70

  involving virus or bacteriophage {(immunoassay for viruses G01N33/56983)} · CPC title

• C12Q1/6818

  involving interaction of two or more labels, e.g. resonant energy transfer · CPC title