Devices comprising bacteriophage PHI6 internal control compositions

Patent metadata

Field	Value
Publication number	US-10422012-B2
Application number	US-201715727116-A
Country	US
Kind code	B2
Filing date	Oct 6, 2017
Priority date	Oct 10, 2016
Publication date	Sep 24, 2019
Grant date	Sep 24, 2019

Abstract

The disclosure provides control compositions used in nucleic acid amplification. Various modifications to the disclosed compositions and methods of using the same, devices, and kits are described.

First claim

What is claimed:

1. A device configured to perform a viral load analysis in a sample, said device comprising
(a) a sample introduction port adapted to receive a sample aliquot;
(b) an internal control compartment comprising a composition comprising bacteriophage Phi6 suspended in a buffer suitable for a nucleic acid amplification reaction at a pH between 7-9, including a chelating agent and a preservative; and
(c) a PCR analysis region comprising one or more additional compartments each configured to conduct one or more steps of said PCR analysis comprising reagent preparation, target enrichment, inhibitor removal, nucleic acid extraction, amplification, and real-time detection.

2. The device of claim 1 wherein said sample comprises plasma, whole blood, urine, fecal material, mucus, and combinations thereof.

3. The device of claim 1 further comprises a swab.

4. The device of claim 1 wherein the sample is suspended in universal transport medium.

5. The device of claim 1 wherein said analysis is an HIV viral load analysis.

6. The device of claim 1 wherein said analysis is a hepatitis or cytomegalovirus viral load analysis.

7. The device of claim 6 wherein said hepatitis is hepatitis B or hepatitis C.

8. The device of claim 1 wherein the composition further comprises one or more additional components comprising BSA, gelatin, poly rA RNA, glycerol, amino acids, trehalose, casein, or polyethylene glycol.

9. The device of claim 1 wherein said buffer is selected from a citrate buffer, a glycine buffer, a tricine buffer, PIPES, and HEPES.

10. The device of claim 1 wherein said bacteriophage Phi6 is present at a concentration of up to 4e5 copies/mL.

11. The device of claim 1 wherein the chelating agent is selected from EDTA and EGTA.

12. The device of claim 1 wherein the preservative is selected from sodium azide and ProClin 300.

13. The device of claim 1 wherein the composition comprises
(a) up to 4e5 cp/mL of Phi6 in 10 mM Tris, pH 8.0, 0.1 mM EDTA, 0.05% Sodium Azide, and 25 mg/mL BSA;
(b) up to 4e5 cp/mL of Phi6 in 10 mM Tris, pH 8.0, 0.1 mM EDTA, 0.05% Sodium Azide, and 1% gelatin;
(c) up to 4e5 cp/mL of Phi6 in 10 mM Tris, pH 8.0, 0.1 mM EDTA, 0.05% Sodium Azide, and 20 mg/L poly rA RNA; or
(d) up to 4e5 cp/mL of Phi6 in 10 mM Tris, pH 8.0, 0.1 mM EDTA, 0.05% Sodium Azide, 20 mg/L poly rA RNA, and 40% glycerol.

14. The device of claim 1 wherein the composition is thermostabile at 45° C. for up to 28 days.

15. The device of claim 1 wherein the composition is thermostabile at 30° C. for up to 110 days.

Assignees

• Roche Molecular Systems Inc

Inventors

• Schoenbrunner Nancy
• Sefah Kwame
• Tian Yu
• Wang Fangnian
• Wang Karen

Classifications

• C12Q1/6851 Primary

  Quantitative amplification · CPC title

• C12Q1/705

  for herpetoviridae, e.g. herpes simplex, varicella zoster · CPC title

• C12Q2600/166

  Oligonucleotides used as internal standards, controls or normalisation probes · CPC title

• C12Q1/707

  non-A, non-B Hepatitis, excluding hepatitis D · CPC title

• C12Q1/703 Primary

  Viruses associated with AIDS · CPC title