Compositions and methods for detecting predisposition to a substance use disorder
View Patent ↗The present invention provides screening kits, compositions, and diagnostic methods for determining whether a subject has a predisposition to, or likelihood of having, a substance use disorder by determining a nucleic acid methylation profile from a biological sample from the subject, wherein a given profile indicates that the subject has a predisposition to a substance use disorder.
1. A Method for detecting that a subject is a user of tobacco comprising:
(a) providing a biological sample from the subject;
(b) contacting DNA from the biological sample with bisulfite under alkaline conditions;
(c) contacting the bisulfite-treated DNA with at least one first oligonucleotide probe at least 8 nucleotides in length that is complementary to a sequence that comprises a CpG dinucleotide at position 373378 of chromosome 5 within the AHRR gene, wherein the at least one first oligonucleotide probe detects either the unmethylated CpG dinucleotide or the methylated CpG dinucleotide, and
(d) detecting either the unmethylated CpG dinucleotide or the methylated CpG dinucleotide,
wherein methylation of the CpG dinucleotide at position 373378 of chromosome 5 is associated with the subject's use of tobacco.
2. The method of claim 1 , wherein the biological sample is peripheral blood.
3. The method of claim 1 , wherein the biological sample is predominantly lymphocytes.
4. The method of claim 1 , further comprising contacting the bisulfite-treated DNA with at least one second oligonucleotide probe at least 8 nucleotides in length that is complementary to the sequence that comprises the CpG dinucleotide at position 373378 of chromosome 5, wherein the at least one second oligonucleotide probe detects either the unmethylated CpG dinucleotide or the methylated CpG dinucleotide that is not detected by the at least one first oligonucleotide probe.
5. The method of claim 4 , further comprising determining the ratio of methylated one or more CpG dinucleotides to unmethylated one or more CpG dinucleotides.
6. The method of claim 1 , further comprising an amplifying step after the contacting step.
7. The method of claim 1 , further comprising a sequencing step after the contacting step.