Comparison of the Methylation Pattern of DNA in Buccal Swabs From Different Time Points
Trial Snapshot
- Phase
- Not Applicable
- Status
- Terminated
- Enrollment
- 20
- Locations
- 1
- Primary Endpoint
- %Methylation (%)
Study Overview
Brief Summary
A buccal swab will be taken using a cotton stick by rubbing it against the buccal mucosa. Samples will be taken from the same participant at different timepoints.
DNA will be extracted from the buccal cells on the swabs using a commercial extraction kit and will be quantified by a nano-drop spectrophotometer.
We will determine global and gene-specific methylation and hydroxymethylation alterations by UPLC/MS/MS. Hydrolyzed DNA into individual nucleosides will be analyzed for the quantitative measurement of 5-methylCytosine and 5-hydroxymethylCytosine on Triple Quadrupole UPLC/MS/MS platform. Secondly, specific methylation in the CpG islands of tumor suppressor and promoter genes, and genes involved in the oxidative stress pathways will be assessed by PCR-pyrosequencing.
Study Design
- Study Type
- Observational
- Observational Model
- Cohort
- Time Perspective
- Prospective
Eligibility Criteria
- Ages
- 18 Years to 35 Years (Adult)
- Sex
- All
- Accepts Healthy Volunteers
- Yes
Inclusion Criteria
- •Caucasian
Exclusion Criteria
- Not provided
Outcomes
Primary Outcomes
%Methylation (%)
Time Frame: 5 weeks
Secondary Outcomes
- %Hydroxymethylation (%)(5 weeks)