# US 2014/0154667 A1 · Solid matrix for one step nucleic acid amplification

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- Number: US 2014/0154667 A1
- Document ID: US20140154667A1
- Status: Published application
- Kind: A1
- Application number: 13/799,174
- Filed: 2013-03-13
- Published: 2014-06-05
- Earliest claimed: 2012-09-13
- Assignees: Ge Healthcare UK Limited
- Applicants: Ge Healthcare UK Limited
- Inventors: KATHRYN L. LAMERTON; CHERYL L. POTTS; SIMON L. STUBBS; MICHAEL K. KENRICK
- Web page: https://patentgrep.com/patent/US20140154667A1

## CPC

- C12N 15/1006: {by means of a solid support carrier, e.g. particles, polymers}
- C12Q 1/6848: characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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

The present invention to relates to methods and kits which can be used to amplify nucleic acids with the advantage of decreasing user time and possible contamination. The PCR reagents are bound to a solid matrix for easy processing and amplification of DNA samples.

## Claims

### Claim 1

A solid matrix for storing and/or amplification of nucleic acids comprising a lysis reagent and a sequestering reagent.

### Claim 2 (depends on 1)

The solid matrix of claim 1, wherein said lysis reagent comprises an anionic surfactant or detergent.

### Claim 3 (depends on 2)

The solid matrix of claim 2, wherein said anionic surfactant is sodium dodecyl sulphate (SDS).

### Claim 4 (depends on 1)

The solid matrix of claim 1, wherein said sequestering agent is a cyclodextrin.

### Claim 5 (depends on 4)

The solid matrix of claim 4, wherein the cyclodextrin is selected from the group consisting of α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin and derivatives thereof.

### Claim 6 (depends on 4)

The solid matrix of claim 4, wherein the cyclodextrin is α-cyclodextrin.

### Claim 7 (depends on 4)

The solid matrix of claim 4, wherein the cyclodextrin is selected from the group consisting of 6-O-α-D-Maltosyl-β cyclodextrin, hydroxyethyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin and 2-hydroxypropyl-β-cyclodextrin.

### Claim 8 (depends on 1)

The solid matrix of claim 1, wherein the solid matrix comprises a glass or silica-based solid phase medium, a plastics-based solid phase medium or a cellulose-based solid phase medium.

### Claim 9 (depends on 8)

The solid matrix of claim 8, wherein the solid phase medium is selected from one of the following: glass, glass fiber, glass microfiber, silica, silica gel, silica oxide, cellulose, nitrocellulose, carboxymethylcellulose, polyester, polyamide, carbohydrate polymers, polypropylene, polytetraflurorethylene, polyvinylidinefluoride, wool or porous ceramics.

### Claim 10 (depends on 1)

The solid matrix of claim 1, wherein the solid matrix is a cellulose based matrix.

### Claim 11 (depends on 10)

The solid matrix of claim 10, wherein said cellulose based matrix is in the form of a pre punched disc.

### Claim 12 (depends on 10)

The solid matrix of claim 10, wherein said cellulose based matrix is in the form of an FTA pre punched disc.

### Claim 13 (depends on 1)

The solid matrix of claim 1, wherein the sequestering reagent and the lysis reagent are separated physically in the matrix.

### Claim 14 (depends on 13)

The solid matrix of claim 13, wherein the lysis reagent is adjacent to the sequestering reagent on said solid matrix.

### Claim 15 (depends on 13)

The solid matrix of claim 13, wherein the sequestering reagent and the lysis reagent are separated by a spacer.

### Claim 16 (depends on 1)

The solid matrix of claim 1, further comprising lyophilised or impregnated or printed Ready to Go PCR reagent for one step nucleic acid amplification wherein the Ready to Go PCR beads comprise of a DNA polymerase, deoxyribonucleotide triphosphate (dNTP), a reaction buffer and at least one primer.

### Claim 17 (depends on 16)

The solid matrix of claim 16, wherein the lysis reagent, sequestrant and PCR regents are physically separated by a spacer.

### Claim 18 (depends on 1)

A method for amplification of nucleic acid using the polymerase chain reaction comprising the steps:
i) contacting the solid matrix of claim 1 with a cellular sample containing nucleic acid;
ii) transferring solid matrix to a reaction vessel; and
iii) incubating the nucleic acid with a PCR reagent mixture for amplification of the nucleic acid.

### Claim 19 (depends on 18)

The method of claim 18, wherein said PCR reagents comprise a DNA polymerase, deoxyribonucleotide triphosphate (dNTP), a reaction buffer and at least one primer.

### Claim 20 (depends on 16)

A method for amplification of nucleic acid using the polymerase chain reaction comprising the steps:
i) contacting the solid matrix of claim 16 with a cellular sample containing nucleic acid;
ii) transferring solid matrix to a reaction vessel; and
iii) incubating the nucleic acid mix with liquid water and forward and reverse primers for amplification of the nucleic acid.

### Claim 21 (depends on 18)

A method of detecting the amplified nucleic acid using a detection system comprising the steps:
i) amplifying the nucleic acids using the method of claim 18; and
ii) detecting the amplified nucleic acid with a computer.

### Claim 22 (depends on 21)

The method of claim 21, wherein the detection system is a PCR imaging system.

### Claim 23 (depends on 18)

The method of claim 18, wherein the cellular sample is selected from a group comprising virus, bacteria and tissue culture cells.

### Claim 24 (depends on 1)

A kit for storing and amplifying the nucleic acid comprising the solid matrix of claim 1 and instructions for use thereof.

### Claim 25 (depends on 16)

A kit for storing and amplifying the nucleic acid comprising the solid matrix of claim 16 and instructions for use thereof.

### Claim 26 (depends on 20)

A method of detecting the amplified nucleic acid using a detection system comprising the steps:
i) amplifying the nucleic acids using the method of claim 20; and
ii) detecting the amplified nucleic acid with a computer.

### Claim 27 (depends on 20)

The method of claim 20, wherein the cellular sample is selected from a group comprising virus, bacteria and tissue culture cells.