Simultaneous collection of DNA and non-nucleic analytes

• US 6,309,827 B1
• Filed: 03/27/1998
• Issued: 10/30/2001
• Est. Priority Date: 03/28/1997
• Status: Expired due to Term

• Current Assignee: Orasure Technologies Incorporated
• Original Assignee: Orasure Technologies Incorporated
• Inventors: Bestwick, Richard K., Goldstein, Andrew S.
• Primary Examiner(s): Carlson, Karen Cochrane
• Assistant Examiner(s): Robinson, Hope A.
• Application Number:
  US09/049,714
• Time in Patent Office: 1,313 Days
• Field of Search: 435/6, 435/70.1, 435/5, 435/7.9, 435/7.25, 435/4, 435/91.2, 436/533, 436/811, 436/17, 436/8, 436/176, 436/63, 436/177, 128/760, 422/56, 525/54.1, 536/22.1, 536/24.33, 536/25.32
• US Class Current: 435/6.11
• CPC Class Codes: C12N 15/1006 by means of a solid support carrier, e.g. particles, polymers
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 1/703 Viruses associated with AIDS
• CPC Class Codes: C12Q 2565/625 being a nucleic acid test strip device, e.g. dipsticks, strips, tapes, CD plates
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: Y10S 436/811 Test for named disease, body condition or organ function
• CPC Class Codes: Y10T 436/10 Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
• CPC Class Codes: Y10T 436/107497 Preparation composition [e.g., lysing or precipitation, etc.]
• CPC Class Codes: Y10T 436/2525 Stabilizing or preserving
• CPC Class Codes: Y10T 436/25375 Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]

Abstract

This invention provides for a rapid and convenient method of simultaneous collection of both genomic and diagnostic information from a single sample on a bibulous pad by differential extraction of the diagnostic information from the genomic information. It is a surprising discovery of this invention that a PCR assay on the contents of the bibulous pad provides results comparable in reliability, specificity, and sensitivity to the best available serum (blood) based assays. The assays of this invention can be used to confirm each other, either by detecting the genomic information leading to the diagnostic information, or by detecting in the genomic information, a predisposition to a disease and confirming the presence of the disease through diagnostic testing.

Claims

1. A method for obtaining and detecting a genomic marker and an extracellular analyte from a single collection step, said method comprising:

(i) collecting a sample onto a bibulous solid support having a fibrous matrix, said sample comprising both (a) whole cells containing a genomic marker and (b) an extracellular analyte;

(ii) selectively obtaining extracellular analyte under conditions in which the genomic marker remains in cells within the fibrous matrix of the solid support;

(iii) obtaining genomic marker from the solid support by lysing the whole cells and freeing genomic marker from the fibrous matrix of the solid support; and

,

(iv) detecting the analyte and genomic marker.

2. A method of claim 1, wherein the sample is a oral mucosal transudate.

3. A method of claim 1, wherein the sample is from a rectal, vaginal or nasal collection.

4. A method of claim 1, wherein the sample is from a blood collection.

8. A method of claim 1, wherein the extracellular analyte is obtained using a solution comprising a non-ionic detergent.

9. The method of claim 8, wherein the solution comprises 0.05-0.5% Tween-20.

10. The method of claim 1, wherein the bibulous solid support comprises an alkali metal compound selected from the group consisting of sodium chloride;

potassium chloride;

magnesium sulfate;

magnesium chloride; and

calcium chloride.

11. The method of claim 10, wherein the bibulous solid support comprises sodium chloride in a concentration such that the concentration in the dry pad is 1-20 mg/cm².

12. The method of claim 8, wherein the solution comprises preservatives selected from the group consisting of chlorhexidine digluconate;

sodium benzoate;

potassium sorbate;

sodium azide;

mercury-containing salts;

sulfate salts of gentamicin, chloramphenicol and streptomycin;

protease inhibitors; and

phenolic compounds.

13. The method of claim 12, wherein the concentration of preservatives is from about 0.001-0.5%.

14. A method of claim 1, wherein the extracellular analyte is selectively extracted from the solid support by pressure, aspiration or centrifugation.

15. A method oc claim 1, wherein the genomic extractions are conducted by impregnating the solid support with a solution that disrupts cell membranes.

16. The method of claim 15, wherein the solution comprises Tris-HCl, pH of about 8.0, in a concentration of about 0.001-0.2 M.

17. The method of claim 15, wherein the solution comprises a chelating agent in a concentration of about 0.05-0.15 M.

18. The method of claim 15, wherein the solution comprises Proteinase K at a concentration of about 50-150 μ

g/mL.

19. A method of claim 15, wherein the solution that disrupts cell membranes comprises an ionic detergent.

20. The method of claim 19, wherein the solution comprises sodium dodecyl sulfate in a concentration of about 0.2-1%.

21. A method of claim 1, wherein the extracellular analyte is detected by an immunoassay.

22. A method of claim 1, wherein the genomic marker is amplified prior to detection by a hybridization probe.

5. A method of obtaining a genomic marker and an extracellular analyte from a single collection step, wherein the extracellular analyte is selected from the group consisting of therapeutic drugs, drugs of abuse, anti-infectious drugs, and disease markers.

6. The method of claim 5, wherein the diagnostic marker is selected from the group consisting of drugs of abuse.

7. The method of claim 6, wherein the diagnostic marker is selected from the group consisting of cannabinoids, cocaine and its metabolites, and methamphetamines and their metabolites.