# US 9,518,303 B2 · Methods and compositions for identifying enterovirus

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 9,518,303 B2
- Document ID: US9518303B2
- Status: Granted
- Kind: B2
- Application number: 14/353,832
- Filed: 2012-10-25
- Published: 2015-10-15
- Granted: 2016-12-13
- Earliest claimed: 2011-10-25
- Assignees: Board of Regents, The University of Texas System
- Applicants: The Board of Regents of the University of Texas System; Richard B. Pyles; Aaron L. Miller
- Inventors: Richard B. Pyles; Aaron L. Miller
- Web page: https://patentgrep.com/patent/US9518303B2

## CPC

- C12Q 1/701: {Specific hybridization probes}
- 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

## Abstract

Certain embodiments of the invention include, but are not limited to PCR primer pairs, sequencing primers, and/or associated thermocycling protocols targeting a region identified by the inventors within the 5′ untranslated region of enteroviruses for the purpose of identifying, subtyping, and/or classifying of virus in samples using nucleic acid sequencing. The sequencing procedures can use nucleic acid templates, such as cDNA or PCR amplicons, as a template for sequencing in medium to high throughput format that is cost effective and easily deployed to other clinical microbiology laboratories.

## Claims

### Claim 1

A method of amplifying enterovirus in a sample comprising:
(a) amplifying a nucleic acid segment from the sample, wherein the nucleic acid segment is an enterovirus 5′UTR corresponding to a nucleic acid segment of rhinovirus A consisting of the nucleic acid sequence of SEQ ID NO:5 to produce an amplicon; and
(b) sequencing the amplicon.

### Claim 2 (depends on 1)

The method of claim 1, wherein more than one enterovirus is present in the sample.

### Claim 3 (depends on 2)

The method of claim 2, wherein a plurality of amplicons are sequenced.

### Claim 4 (depends on 1)

The method of claim 1, wherein the sequencing is by pyrosequencing.

### Claim 5 (depends on 1)

The method of claim 1, wherein the sample is a blood, urine, lymph, sputum, saliva, or tissue sample.

### Claim 6 (depends on 1)

The method of claim 1, wherein the amplifying step uses amplification primers comprising a nucleotide sequence of SEQ ID NO:1 and SEQ ID NO:2.

### Claim 7 (depends on 1)

The method of claim 1, wherein the sequencing step uses a sequencing primer comprising a nucleic acid sequence of SEQ ID NO:3.