Am J Epidemiol 2004; 160:77-84.
Copyright © 2004 by the Johns
Hopkins Bloomberg School of Public Health
PRACTICE OF EPIDEMIOLOGY |
Optimizing Buccal Cell DNA Yields in Mothers and Infants for Human Leukocyte Antigen Genotyping
1 Department of Epidemiology, University of Iowa College of Public Health, Iowa City, IA.
2 Veterans Affairs Medical Center, Iowa City, IA.
3 Center for Perinatal, Pediatric and Environmental Epidemiology, Department of Epidemiology and Public Health, Yale School of Medicine, New Haven, CT.
Buccal cells provide a convenient source of DNA for epidemiologic studies. Mouthwash rinses yield a higher quality and quantity of DNA than cytobrushes but are not practical for collection from infants. Although cytobrushes yield sufficient DNA for most genotyping analyses, human leukocyte antigen (HLA) analysis can require 1,000-fold more DNA. In Iowa City, Iowa, in 2002, the authors tested two cytobrush collection methods to optimize total DNA yield and purity for HLA genotyping in mothers and infants: 1) brushing the left and right inner cheeks (standard method) and 2) brushing the upper and lower "gutters", that is, the space between the gums and the inner lips/cheeks along the front and sides of the mouth (test method). Storage and mailing experiments were performed to define conditions for optimizing DNA yield and purity. Mothers’ gutter samples yielded significantly higher total amounts of DNA (mean yield = 15.0 µg/two brushes) than cheek samples (mean yield = 7.6 µg/two brushes) (paired t test: p < 0.001), while DNA yields from cheek and gutter collections from infants were equivalent. Cytobrushes stored and/or mailed in paper envelopes yielded significantly more and higher-purity DNA than brushes in plastic bags or tubes. Cytobrush sampling of the mouth’s gutter areas can enhance DNA yield in mothers but not in young infants. DNA yields can be further optimized by controlling mailing and storage conditions.
DNA; HLA antigens; mouth mucosa; specimen handling
Abbreviations: Abbreviations: HLA, human leukocyte antigen; SD, standard deviation.
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