Generic matrix for control nucleic acids

Patent metadata

Field	Value
Publication number	US-9920354-B2
Application number	US-201113328280-A
Country	US
Kind code	B2
Filing date	Dec 16, 2011
Priority date	Dec 17, 2010
Publication date	Mar 20, 2018
Grant date	Mar 20, 2018

Abstract

The present invention is in the field of in-vitro diagnostics. Within this field, it provides the amplification of at least a first target nucleic acid that may be present in at least one fluid sample using an internal control nucleic acid for qualitative and/or quantitative purposes and at least one external control nucleic acid in an aqueous buffer. It further provides an analytical system comprising an internal control nucleic acid for qualitative and/or quantitative purposes and at least one external control nucleic acid in an aqueous buffer.

First claim

What is claimed is:

1. A process for controlled amplification of a target nucleic acid that may be present in a fluid sample, comprising:
(a) providing in a vessel an internal control nucleic acid in the fluid sample,
(b) providing a solid support material to the vessel for a period of time and under conditions sufficient to permit nucleic acids comprising the target nucleic acid and the internal control nucleic acid to be immobilized on the solid support material,
(c) isolating the solid support material from other material present in the vessel in a separation station,
(d) purifying the nucleic acids in the separation station and washing the solid support material one or more times with a wash buffer,
(e) providing (i) in a first reaction vessel the purified nucleic acids of step (d), wherein the purified nucleic acids comprise the internal control nucleic acid and the target nucleic acid derived from the fluid sample, and (ii) in a second reaction vessel an external control nucleic acid in an aqueous buffer, wherein the aqueous buffer lacks a fluid sample,
(f) adding identical amplification reagents to each of the first and second reaction vessels,
(g) incubating the first and second reaction vessels under identical conditions sufficient for amplification reactions to occur, and
(h) detecting results of the amplification reactions, wherein the results of the amplification reactions are indicative of the presence or absence of amplification of the target nucleic acid, the internal control nucleic acid and the external control nucleic acid
wherein the aqueous buffer has a pH between 6.0 and 12.0 and comprises: 1-100 mM Tris, 0.01-1 mM EDTA, 0.005-0.5% (w/v) Sodium Azide, and 1-200 mg/l Poly(rA) RNA.

2. The process of claim 1 , wherein the aqueous buffer has a pH of about 8 and comprises: 10 mM Tris, 0.1 mM EDTA, 0.05% (w/v) Sodium Azide, and 20 mg/l Poly(rA) RNA.

3. The process of claim 1 , wherein a negative control is provided in a third reaction vessel and is subjected to steps (b) through (h).

4. The process of claim 1 , wherein the fluid sample is a clinical sample.

5. The process of claim 1 , further comprising: (i) determining the quantity of the target nucleic acid.

6. The process of claim 1 , wherein the fluid sample comprises blood.

7. The process of claim 1 , wherein the fluid sample comprises blood plasma.

8. The process of claim 1 , wherein the aqueous buffer is free of biological buffers derived from blood.

9. The process of claim 1 , wherein the aqueous buffer is free of biological buffers derived from blood plasma.

10. The process of claim 1 , wherein the second reaction vessel comprises more than one external control nucleic acid.

Assignees

• Eickhoff Meike
• Russmann Eberhard
• Woelfelschneider Andreas
• Zimmermann Dirk
• Roche Molecular Systems Inc

Inventors

• Eickhoff Meike
• Russmann Eberhard
• Woelfelschneider Andreas
• Zimmermann Dirk

Classifications

• C12Q2545/101

  with an internal standard/control · CPC title

• C12Q1/703

  Viruses associated with AIDS · CPC title

• C12Q2537/143

  Multiplexing, i.e. use of multiple primers or probes in a single reaction, usually for simultaneously analyse of multiple analysis · CPC title

• C12Q1/6806 Primary

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• C12Q2545/113

  with an external standard/control, i.e. control reaction is separated from the test/target reaction · CPC title