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Buccal swabs and treated cards: methodological considerations for molecular epidemiologic studies examining pediatric populations.
- Source :
-
American journal of epidemiology [Am J Epidemiol] 2008 May 15; Vol. 167 (10), pp. 1260-7. Date of Electronic Publication: 2008 Mar 07. - Publication Year :
- 2008
-
Abstract
- Self-collection of buccal cells provides a noninvasive method for obtaining biologic samples for genetic analyses in pediatric studies. Nevertheless, low yields, microbial contamination, and degradation of buccal samples present challenges for epidemiologic studies incorporating genetic investigations. The aims of this study were to compare the quality and yield of DNA extracted from buccal specimens with BuccalAmp swabs (Epicenter BioTechnologies, Madison, Wisconsin) or FTA cards (Whatman, Inc., Clifton, New Jersey) and to investigate the use of whole-genome amplification (WGA) for increasing DNA yields for single nucleotide polymorphism analyses. Buccal specimens were collected from 55 children with acute lymphoblastic leukemia and 52 control children without acute lymphoblastic leukemia in New South Wales, Australia, in 2003-2004. Real-time polymerase chain reaction was used to evaluate polymorphisms in the genes encoding the cytochrome p450 enzyme CYP3A4 (CYP3A4 A392G, also known as CYP3A4*1B) and the steroid xenobiotic receptor (SXR C25385T). Results showed that DNA could be isolated from buccal specimens collected by use of both methods and that yields could be substantially improved with WGA without introducing genotyping error. However, DNA quality was poorer in samples collected by BuccalAmp swabs, and the presence of polymerase chain reaction inhibitors in these samples reduced the sensitivity of quantitative real-time PCR analysis. These findings show that different methods for collecting buccal samples impact on the downstream success of genetic investigations and influence DNA quality after WGA.
- Subjects :
- Cheek
Child
Female
Genotype
Humans
Male
Nucleic Acid Amplification Techniques instrumentation
Paper
Pediatrics methods
Polymerase Chain Reaction
Specimen Handling methods
DNA analysis
Molecular Epidemiology methods
Mouth Mucosa cytology
Nucleic Acid Amplification Techniques methods
Polymorphism, Single Nucleotide
Sequence Analysis, DNA methods
Subjects
Details
- Language :
- English
- ISSN :
- 1476-6256
- Volume :
- 167
- Issue :
- 10
- Database :
- MEDLINE
- Journal :
- American journal of epidemiology
- Publication Type :
- Academic Journal
- Accession number :
- 18326872
- Full Text :
- https://doi.org/10.1093/aje/kwn012