Methods and kits for obtaining and analyzing skin samples for the detection of nucleic acids

• US 6,949,338 B2
• Filed: 08/17/1999
• Issued: 09/27/2005
• Est. Priority Date: 08/18/1998
• Status: Expired due to Term

• Current Assignee: DermTech International
• Original Assignee: DermTech International
• Inventors: Rheins, Lawrence A., Morhenn, Vera B.
• Primary Examiner(s): Spector, Lorraine
• Application Number:
  US09/375,609
• Publication Number:
  US 20020197604A1
• Time in Patent Office: 2,233 Days
• Field of Search: 204/451, 435/6
• US Class Current: 435/6.11
• CPC Class Codes: A61K 49/0006 Skin tests, e.g. intradermal testing, test strips, delayed hypersensitivity
• 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 2600/158 Expression markers

Abstract

Disclosed is a method for removing polynucleotide from the skin. This polynucleotide can be used to detect dermatitis and distinguish an irritant reaction from an allergic reaction by characterizing the polynucleotide according to the polypeptide which it encodes. Additionally, provided are methods for non-invasive isolation of samples from the skin as well as kits for use in the methods provided herein.

Claims

1. A method for quantitating relative expression of a ribonucleic acid (RNA) from a skin sample, comprising:

(a) applying an adhesive tape to the skin, under conditions allowing for isolation of a skin sample adhering to the adhesive tape;

(b) detecting the RNA from the skin sample; and

(c) comparing the level of the RNA in the skin sample to a control sample, thereby quantitating relative expression of the RNA.

3. The method of claim 1, wherein the detecting comprises detecting mRNA.

4. The method of claim 3, wherein the detecting comprises detecting an RNA that encodes an inflammatory mediator.

5. The method of claim 1, wherein the detecting comprises detecting an RNA that encodes a cytokine.

6. The method of claim 1, wherein the detecting comprises detecting an RNA that encodes an interleukin.

7. The method of claim 1, wherein the detecting comprises or detecting an RNA that encodes interleukin-1 (IL-1), interleukin-2 (IL-2), interleukin-3 (IL-3), interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), interleukin-12 (IL-12), interleukin-13 (IL- 13), granulocyte macrophage colony stimulating factor (GM-CSF), or an interferon, or any combination thereof.

8. The method of claim 1, wherein the adhesive tape is applied to skin from a subject afflicted with a disease, disorder, or inflammatory reaction.

9. The method of claim 8, wherein the adhesive tape is applied to skin from a subject afflicted with dermatitis.

10. The method of claim 1, wherein the method further comprises contacting the skin with an external agent that causes dermatitis before applying the adhesive tape to the skin.

11. The method of claim 1, wherein the subject is a mammal.

12. The method of claim 1, wherein the subject is a human.

13. The method of claim 1, wherein the subject is a non-human.

2. The method of claim 2, wherein the skin sample comprises stratum corneum cells and cells associated with the stratum corneum which are removed by application and removal of the adhesive tape.