# US 10,308,989 B2 · Aptamers with binding affinity to norovirus

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- Number: US 10,308,989 B2
- Document ID: US10308989B2
- Status: Granted
- Kind: B2
- Application number: 15/317,002
- Filed: 2015-06-12
- Published: 2017-04-27
- Granted: 2019-06-04
- Earliest claimed: 2014-06-13
- Assignees: North Carolina State University
- Applicants: North Carolina State University
- Inventors: Lee-Ann Jaykus; Blanca Irene Escudero-Abarca; Matthew D. Moore; Helen Rawsthorne
- Web page: https://patentgrep.com/patent/US10308989B2

## CPC

- C12Q 1/70: involving virus or bacteriophage {(immunoassay for viruses G01N33/56983)}
- B82Y 30/00: Nanotechnology for materials or surface science, e.g. nanocomposites
- C12N 15/115: Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith {; Nucleic acids binding to non-nucleic acids, e.g. aptamers}
- C12Q 1/6834: Enzymatic or biochemical coupling of nucleic acids to a solid phase
- G01N 33/5308: {for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites}
- G01N 33/56983: {Viruses}
- C12N 2310/16: Aptamers
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- B82Y: SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

The instant disclosure provides norovirus binding aptamers, compositions comprising such aptamers, and methods of using and producing such aptamers. The aptamers are useful, for example, for detecting the presence of norovirus in test samples, for capturing and/or concentrating norovirus from test samples, for evaluating the efficacy of therapeutic agents in patients diagnosed with a norovirus infection, and for evaluating the efficacy of norovirus vaccines.

## Claims

### Claim 1

A method of detecting the presence of at least one norovirus strain in a test sample, the method comprising:
(a) contacting said test sample with a norovirus-binding aptamer comprising:
(i) a norovirus-binding motif, wherein said norovirus-binding motif comprises one or more of SEQ ID NOS: 98-100, 162-164, and 112-120; and/or
(ii) the nucleic acid sequence of SEQ ID NO: 25 any one of SEQ ID NOS: 1 78 and 176 181 or variants thereof having at least 90% sequence identity and having norovirus-binding activity; and
(b) detecting the presence of said norovirus-binding aptamer bound to norovirus in said test sample, wherein detection of bound aptamer indicates the presence of at least one norovirus strain.

### Claim 2 (depends on 1)

The method of claim 1, further comprising removing unbound norovirus-binding aptamer prior to detecting the presence of said norovirus-binding aptamer bound to norovirus in said test sample.

### Claim 3 (depends on 1)

The method of claim 1, further comprising comparing the presence of said norovirus-binding aptamer bound to norovirus in said test sample with the presence of said norovirus-binding aptamer bound to norovirus in a control sample, whereby increased presence of said norovirus-binding aptamer bound to norovirus in said test sample relative to said control sample indicates the presence of at least one norovirus strain in said test sample.

### Claim 4 (depends on 1)

The method of claim 1, wherein said test sample is a clinical sample.

### Claim 5 (depends on 1)

The method of claim 1, wherein said test sample is an environmental sample or a food sample.

### Claim 6 (depends on 1)

The method of claim 1, wherein the detecting step comprises amplifying the bound aptamer.

### Claim 7 (depends on 1)

The method of claim 1, further comprising capturing said at least one norovirus strain from said test sample by substantially separating the aptamer-bound norovirus from the remainder of said test sample.

### Claim 8 (depends on 1)

The method of claim 1, wherein said norovirus-binding aptamer preferentially binds to norovirus strains from genogroup I; genogroup I, genotype 1; genogroup II; genogroup II, genotype 2; and/or genogroup II, genotype 4.

### Claim 9 (depends on 1)

The method of claim 1, wherein said norovirus-binding aptamer preferentially binds to an epitope within the VPg protein, the VP1 protein, the P domain of the VP1 protein, the PI subdomain of the VP1 protein, the P2 subdomain of the VP1 protein, the S domain of the VP1 protein, and/or the VP2 protein of said at least one norovirus strain.

### Claim 10 (depends on 1)

The method of claim 1, wherein said norovirus-binding aptamer is a single-stranded DNA aptamer.

### Claim 11 (depends on 1)

The method of claim 1, wherein said norovirus-binding aptamer comprises the nucleic acid sequence of SMV-19 (SEQ ID NO: 25).

### Claim 12 (depends on 1)

The method of claim 1, wherein said norovirus-binding motif comprises one or more of SEQ ID NOS: 98-100, 112-120, and 162-164.

### Claim 13 (depends on 1)

The method of claim 1, wherein said norovirus-binding aptamer is tethered to a solid support or immobilized on a magnetic bead.

### Claim 14 (depends on 1)

The method of claim 1, wherein said norovirus-binding aptamer further comprises a label.

### Claim 15 (depends on 1)

The method of claim 1, wherein said norovirus-binding aptamer comprises an aptamer mixture comprising a first aptamer and at least one different aptamer.