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.
Castle
Philip E
PE
Division of Cancer Epidemiology and Genetics,National Cancer Institute, Bethesda, MD 20892, USA. castlep@mail.nih.gov
Garcia-Closas
Montserrat
M
Franklin
Tracie
T
Chanock
Stephen
S
Puri
Vinita
V
Welch
Robert
R
Rothman
Nathaniel
N
Vaught
Jim
J
D023482:Bioterrorism / Q000517:prevention & control; D004247:DNA / Q000528:radiation effects*; D006801:Humans; D009061:Mouth Mucosa / Q000166:cytology / Q000528:radiation effects*; D020641:Polymorphism, Single Nucleotide; D011177:Postal Service; D013242:Sterilization / Q000379:methods; D020080:Tandem Repeat Sequences
D004247:DNA