Effects of electron-beam irradiation on buccal-cell DNA.

Buccal cells were collected from 29 participants, by use of mouthwash rinses, and were split into equal aliquots, with one aliquot irradiated by electron-beam (E-beam) irradiation equivalent to the sterilizing dosage used by the U.S. Postal Service and the other left untreated. Aliquots were extracted and tested for DNA yields (e.g., TaqMan assay for quantifying human genomic DNA), genomic integrity, and amplification-based analysis of genetic variants (e.g., single-nucleotide polymorphisms [SNPs] and single tandem repeats [STRs]). Irradiated aliquots had lower median DNA yields (3.7 microg/aliquot) than untreated aliquots (7.6 microg/aliquot) (P<.0005) and were more likely to have smaller maximum DNA fragment size, on the basis of genomic integrity gels, than untreated aliquots (P<.0005). Irradiated aliquots showed poorer PCR amplification of a 989-bp beta-globin target (97% for weak amplification and 3% for no amplification) than untreated aliquots (7% for weak amplification and 0% for no amplification) (P<.0005), but 536-bp and 268-bp beta-globin targets were amplified from all aliquots. There was no detectable irradiation effect on SNP assays, but there was a significant trend for decreased detection of longer STRs (P=.01) in irradiated versus untreated aliquots. We conclude that E-beam irradiation reduced the yield and quality of buccal-cell specimens, and, although irradiated buccal-cell specimens may retain sufficient DNA integrity for some amplified analyses of many common genomic targets, assays that target longer DNA fragments (>989 bp) or require whole-genome amplification may be compromised.

[1]  N. Oldroyd,et al.  Rapid quantification of DNA samples extracted from buccal scrapes prior to DNA profiling. , 1997, BioTechniques.

[2]  A M Nomura,et al.  Feasibility of collecting buccal cell DNA by mail in a cohort study. , 2001, Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology.

[3]  David Hanson ZAPPING THE MAIL , 2002 .

[4]  R. Williamson,et al.  SIMPLE NON-INVASIVE METHOD TO OBTAIN DNA FOR GENE ANALYSIS , 1988, The Lancet.

[5]  J. Gohagan,et al.  The Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial of the National Cancer Institute: history, organization, and status. , 2000, Controlled clinical trials.

[6]  T. Rebbeck,et al.  Collection of genomic DNA by buccal swabs for polymerase chain reaction-based biomarker assays. , 1999, Environmental health perspectives.

[7]  K. Buetow,et al.  Collection of genomic DNA from adults in epidemiological studies by buccal cytobrush and mouthwash. , 2001, Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology.

[8]  E. Buse,et al.  Performance evaluation of two multiplexes used in fluorescent short tandem repeat DNA analysis. , 2003, Journal of forensic sciences.

[9]  C. Wheeler,et al.  Sample preparation and PCR amplification from paraffin-embedded tissues. , 1994, PCR methods and applications.

[10]  M. Manos,et al.  19 – SAMPLE PREPARATION FROM PARAFFIN-EMBEDDED TISSUES , 1990 .

[11]  L. Le Marchand,et al.  A simple mouthwash method for obtaining genomic DNA in molecular epidemiological studies. , 1998, Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology.

[12]  B. Kross,et al.  A model for predicting the frequency of high pesticide exposure events in the Agricultural Health Study. , 2000, Environmental research.