Diagnosis of melanoma and solar lentigo by nucleic acid analysis
The present invention provides methods for diagnosing melanoma and/or solar lentigo in a subject by analyzing nucleic acid molecules obtained from the subject. The present invention also provides methods for distinguishing melanoma from solar lentigo and/or dysplastic nevi and/or normal pigmented skin. The methods include analyzing expression or mutations in epidermal samples, of one or more skin markers. The methods can include the use of a microarray to analyze gene or protein profiles from a sample.
1. A method of detecting expression levels of preferentially expressed antigen in melanoma (PRAME) and C6orf218 in a subject, the method comprising:
a) obtaining a biological sample of a skin lesion suspected of comprising melanoma from the subject with an adhesive tape, wherein the biological sample comprises or is suspected of comprising a nucleic acid molecule expressed from PRAME and a nucleic acid molecule expressed from C6orf218; and
b) detecting the expression level of PRAME and C6orf218 to determine whether the expression levels of PRAME and C6orf218 in the biological sample are at levels that are above expression levels of PRAME and C6orf218 in a non-melanoma sample by a hybridization assay, whereby the presence of a nucleic acid molecule expressed from PRAME and a nucleic acid molecule expressed from C6orf218 in the biological sample in amounts that are greater than the presence of PRAME and C6orf218 in a non-melanoma sample is indicative of melanoma in the biological sample.
2. The method of claim 1 , provided that determining an expression level of the nucleic acid molecule expressed from PRAME and the nucleic acid molecule expressed from C6orf218, wherein the PRAME and C6orf218 nucleic acid molecules each comprise an RNA molecule, comprises generating a cDNA product from each of the PRAME and C6orf218 RNA molecules, and amplifying the cDNA product by real-time quantitative polymerase chain reaction (PCR).
3. The method of claim 1 , wherein the adhesive tape comprises a polyurethane film.
4. The method of claim 3 , provided that about one to about ten adhesive tapes or one to ten applications of the tape are applied to and removed from the skin lesion.
5. The method of claim 1 , wherein the adhesive tape is a pliable tape comprising rubber adhesive.
6. The method of claim 1 , provided that determining an expression level of the nucleic acid molecule expressed from PRAME and the nucleic acid molecule expressed from C6orf218 comprises microarray analysis or a direct sequencing method.
7. The method of claim 1 , wherein the expression level of the corresponding nucleic acid molecule expressed from PRAME and the nucleic acid molecule expressed from C6orf218 in the non-melanoma sample is contained within a database.
8. The method of claim 1 , wherein the hybridization assay comprises application of a detectably labeled probe that hybridizes to a nucleic acid molecule expressed from PRAME and a detectably labeled probe that hybridizes to a nucleic acid molecule expressed from C6orf218.
9. The method of claim 1 , wherein the biological sample comprises cells from the stratum corneum.
10. A method of detecting expression levels of preferentially expressed antigen in melanoma (PRAME) and C6orf218 in a subject, the method comprising:
a) obtaining a biological sample of a skin lesion suspected of comprising melanoma from the subject, wherein the biological sample comprises or is suspected of comprising a nucleic acid molecule expressed from PRAME and a nucleic acid molecule expressed from C6orf218; and
b) detecting the expression level of PRAME and C6orf218 with a detectably labeled probe that hybridizes to a nucleic acid molecule expressed from PRAME and a detectably labeled probe that hybridizes to a nucleic acid molecule expressed from C6orf218 to determine whether the biological sample has elevated expression levels of PRAME and C6orf218 compared to the expression levels of PRAME and C6orf218 in a non-melanoma sample.
11. The method of claim 10 , provided that determining an expression level of the nucleic acid molecule expressed from PRAME and the nucleic acid molecule expressed from C6orf218, wherein the PRAME and C6orf218 nucleic acid molecules each comprise an RNA molecule, comprises generating a cDNA product from each of the PRAME and C6orf218 RNA molecules, and amplifying the cDNA product by real-time quantitative polymerase chain reaction (PCR).
12. The method of claim 10 , wherein the biological sample is obtained with an adhesive tape.
13. The method of claim 12 , wherein the adhesive tape comprises a polyurethane film.
14. The method of claim 13 , provided that about one to about ten adhesive tapes or one to ten applications of the tape are applied to and removed from the skin lesion.
15. The method of claim 12 , wherein the adhesive tape is a pliable tape comprising rubber adhesive.
16. The method of claim 10 , provided that determining an expression level of the nucleic acid molecule expressed from PRAME and the nucleic acid molecule expressed from C6orf218 comprises microarray analysis or a direct sequencing method.
17. The method of claim 10 , wherein the expression level of the corresponding nucleic acid molecule expressed from PRAME and the nucleic acid molecule expressed from C6orf218 in the non-melanoma sample is contained within a database.
18. The method of claim 10 , wherein the biological sample comprises cells from the stratum corneum.