NON-INVASIVE SKIN-BASED DETECTION METHODS
Disclosed herein are analytical methods and compositions for detecting expression level and mutational change in an individual in need thereof, which profiles RNA, genomic DNA, and/or microbial DNA. Also described herein include diagnostic methods which are based on the changes of expression levels and mutational change of RNA, genomic DNA, and/or microbial DNA.
1 . A method of preparing a microbiome sample for analysis, comprising:
(a) providing one or more adhesive patches, wherein the one or more adhesive patches were applied to a skin surface;
(b) isolating genetic material from the one or more adhesive patches; and
(c) identifying one or more RNA or DNA biomarkers present in the genetic material, wherein the one or more RNA or DNA biomarkers are from a microbe, thereby preparing a microbiome sample for analysis.
2 . The method of claim 1 , wherein the one or more RNA or DNA biomarkers are from a plurality of microbes.
3 . The method of claim 1 , wherein the one or more adhesive patches comprises at least 4, 6, 7, 8, or more patches.
4 . The method of claim 1 , wherein the one or more adhesive patches were applied to more than one skin surface.
5 . The method of claim 1 , wherein the genetic material is RNA, DNA, or RNA and DNA.
6 . The method of claim 1 , wherein identifying the one or more DNA or RNA biomarkers comprises using Sanger sequencing, next generation sequencing, single-molecule real-time sequencing, Polony sequencing, sequencing by synthesis, sequencing by ligation, reversible terminator sequencing, proton detection sequencing, ion semiconductor sequencing, nanopore sequencing, electronic sequencing, pyrosequencing, Maxam-Gilbert sequencing, chain termination sequencing, or +S sequencing.
7 . The method of claim 1 , wherein the genetic material comprises pathogenic nucleic acids, bacterial nucleic acids, viral nucleic acids, fungal nucleic acids, parasitic nucleic acids, or any combination thereof.
8 . The method of claim 1 , wherein the microbe comprises a bacterium or a fungus.
9 . The method of claim 8 , wherein the microbe comprises a bacterium, and the bacterium is of the genus Actinomycetales, Anaerococcus, Bacillales, Bifidobacterium, Enhydrobacter, Finegoldia, Carnobacterium, Coryneobacterium, Lactobacillus, Lactococcus, Leunconostoc, Macrooccus, Micrococcineae, Oenococcus, Pediococcus, Peptoniphilus, Propionibacterium, Salinicoccus, Sphingomonas, Staphylococcus, Strepococcus, Tetragenoccus, or Weissella.
10 . The method of claim 8 , wherein the microbe comprises a fungus, and the fungus is of the genus Malassezia, Pityrosporum, Aspergillus, Candida, Cryptococcus, Rhodotorula, or Epicoccum.
11 . The method of claim 1 , wherein identifying comprises determining expression levels of one or more RNA.
12 . The method of claim 1 , wherein identifying comprises determining a presence or absence of a DNA mutation in at least one gene of interest.
13 . A method of preparing a skin sample for analysis, comprising:
providing a non-invasive skin sample, wherein the skin sample was obtained from a subject using one or more adhesive patches;
extracting nucleic acids from the skin sample, wherein the nucleic acids comprise non-microbial nucleic acids and microbial nucleic acids;
detecting an expression level of one or more target genes in the non-microbial nucleic acids from the skin sample; and
detecting a microbiome profile from the microbial nucleic acids.
14 . The method of claim 13 , wherein the non-microbial nucleic acids comprise genomic DNA, RNA, or both genomic DNA and RNA.
15 . The method of claim 13 , wherein detecting a microbiome profile comprises identifying a 16S sequence from the microbial nucleic acids or measuring expression levels of one or more microbial genes.
16 . The method of claim 13 , wherein the microbiome profile is determined by quantification of one or more of: a cell count of one or more microbes, a total cell count of all detected microbes, and a total cell count of all human cells.
17 . The method of claim 13 , wherein detecting the expression level of one or more target genes or detecting the microbiome profile comprises at least one amplification step.
18 . The method of claim 17 , wherein the at least one amplification step comprises quantitative PCR (qPCR), self-sustained sequence replication, transcriptional amplification system, Q-Beta Replicase, or rolling circle replication.
19 . The method of claim 13 , wherein detecting an expression level of one or more target genes or detecting a microbiome profile comprises using qPCR, microarray, or sequencing.
20 . The method of claim 13 , wherein the method further comprises identifying a skin condition when the expression level of one or more target genes is increased or decreased by at least about 5% compared to a control.
21 . The method of claim 13 , wherein the method further comprises identifying a skin condition when the expression level of one or more target genes is increased or decreased by at least about 10% compared to a control.
22 . The method of claim 13 , wherein the skin sample was obtained from the face or forehead.
23 . The method of claim 13 , wherein the method further comprises identifying, predicting, or treating a disease or condition from the expression level of one or more target genes in the non-microbial sample and the microbiome profile.
24 . The method of claim 23 , wherein the disease or condition is a skin disease selected from the group consisting of: psoriasis, atopic dermatitis, seborrhoeic dermatitis, and acne.
25 . A skin sample collection kit for capturing genetic and/or microbial data from a subject, comprising:
at least one adhesive tape comprising a flexible backing film, an adhesive matrix for non-invasively collecting a skin microbe sample from a subject, and a peelable release sheet for protecting the adhesive matrix prior to skin sample collection;
a storage area for receiving the adhesive tape after skin microbe sample collection; and
a unique barcode associated with the skin sample.