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Buccal Swabs and Treated Cards - Methodological Considerations For Molecular Epidemiologic Studies Examining Pediatric Populations. Am. J. Epidemiol.-2008-Beckett-1260-7
Buccal Swabs and Treated Cards - Methodological Considerations For Molecular Epidemiologic Studies Examining Pediatric Populations. Am. J. Epidemiol.-2008-Beckett-1260-7
Buccal Swabs and Treated Cards - Methodological Considerations For Molecular Epidemiologic Studies Examining Pediatric Populations. Am. J. Epidemiol.-2008-Beckett-1260-7
The Author 2008. Published by the Johns Hopkins Bloomberg School of Public Health.
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Practice of Epidemiology
Buccal Swabs and Treated Cards: Methodological Considerations for Molecular
Epidemiologic Studies Examining Pediatric Populations
Childrens Cancer Institute Australia for Medical Research, Randwick, New South Wales, Australia.
Department of Hematology and Oncology, Starship Childrens Hospital, Auckland, New Zealand.
3
Department of Oncology, The Childrens Hospital at Westmead, Westmead, New South Wales, Australia.
4
Center for Childrens Cancer and Blood Disorders, Sydney Childrens Hospital, Randwick, New South Wales, Australia.
5
Telethon Institute for Child Health Research, Center for Child Health Research, University of Western Australia, Perth, Western
Australia, Australia.
2
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 wholegenome amplication (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 20032004. 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 ndings show that different methods for collecting buccal
samples impact on the downstream success of genetic investigations and inuence DNA quality after WGA.
DNA; epidemiologic methods; epidemiology, molecular; genome; mouth mucosa; pediatrics
Abbreviations: MDA, multiple displacement amplication; PCR, polymerase chain reaction; SNP, single nucleotide
polymorphism; WGA, whole-genome amplication.
Correspondence to Dr. Lesley Ashton, Molecular Epidemiology Group, Childrens Cancer Institute Australia for Medical Research, P.O. Box 81,
Randwick, NSW 2031, Australia (e-mail: lashton@ccia.unsw.edu.au).
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Am J Epidemiol 2008;167:12601267
Received for publication June 26, 2007; accepted for publication January 14, 2008.
Am J Epidemiol 2008;167:12601267
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TABLE 1. DNA yields before and after whole-genome amplication in cases and
controls who returned both BuccalAmp swabs* and FTA cards*, New South Wales,
Australia, 20032004
Cases (n 31)
DNA sample
Median DNA
yields
(ng/100 ll)
Controls (n 52)
Interquartile
range
Median DNA
yields
(ng/100 ll)
Interquartile
range
p valuey
BuccalAmp solutions
Before WGAz
After WGA
34.1123.5
26.3
6.169.0
0.007
350
79.1
120.0596.5
78.8
8.8297.0
0.002
240
116.5424.5
134.3371.0
0.443
FTA cards
After WGA
257
FIGURE 2. Correlation between age at collection and the BuccalAmp DNA yields obtained from 79 children, New South Wales,
Australia, 20032004. BuccalAmp (Epicenter BioTechnologies,
Madison, Wisconsin).
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10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
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Am J Epidemiol 2008;167:12601267
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