Integrated nucleic acid analysis

• US 20060014185A1
• Filed: 06/29/2005
• Published: 01/19/2006
• Est. Priority Date: 06/29/2004
• Status: Active Grant

• Current Assignee: Wallac Oy (Revvity, Inc.)
• Original Assignee: Wallac Oy (Revvity, Inc.)
• Inventors: Ylikoski, Alice, Ollikka, Pia
• Granted Patent: US 7,776,530 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2523/308 Adsorption or desorption

Abstract

The present invention relates to an integrated method of nucleic acid analysis, and more particularly to a simplified sample pre-treatment, which renders the method more easily automated, where the sample is provided on or applied onto a solid matrix and the subsequent amplification and detection steps are performed in one single, sealed reaction vial without removing the matrix.

Claims

1. A method for analyzing a target nucleic acid in a biological sample, comprising the following steps

i) providing an absorbent matrix in a reaction vial;

ii) providing a biological sample containing said target nucleic acids on, or applying it onto, said absorbent matrix;

iii) providing in said reaction vial a reagent mixture for amplifying said nucleic acids;

iv) providing in said reaction vial a detection reagent mixture;

v) sealing said reaction vial;

vi) performing an amplification reaction; and

vii) detecting the amplified nucleic acids, wherein

steps vi) and vii) are performed in said vial without removing said matrix from the vial, and wherein the detection step is performed in closed-tubes, without opening the seals.

2. The method according to claim 1, wherein the sample is provided on the absorbent matrix, and at least a portion of said matrix is provided into the vial together with said sample.

3. The method according to claim 2, wherein the sample is applied onto the matrix immediately prior to providing said matrix into the reaction vial, without drying and/or storage.

4. The method of claims 1, wherein the matrix is provided in the reaction vial prior to step ii) and the sample is applied onto said matrix contained in said vial.

5. The method according to claims 1, wherein the matrix is an absorbent material that does not irreversibly bind nucleic acids.

6. The method according to claim 5, wherein the matrix comprises a protein binding and/or a protein denaturing agent.

7. The method according to claim 5, wherein the matrix comprises a chaotropic agent.

8. The method according to claims 1, wherein at least part of the amplification and detection reagent mixtures are contained in the reaction vial in dried form, said method including an optional further step of adding water prior to step v).

9. The method according to claims 1, wherein the amplification is performed by PCR.

10. The method according to claims 1, wherein the detection step is performed by hybridization of the amplified sample to a fluorescently labelled probe.

11. A kit for use in a method according to claims 1, comprising at least one reaction vial comprising an absorbent matrix;

a reagent mixture for amplifying said nucleic acids; and

a detection reagent mixture for detecting said amplified nucleic acids;

wherein at least part of said reagent and detection mixtures are provided in dried form.

12. A kit according to claim 11, wherein all reagents are provided in dried form in said reaction vial.