Buccal cell DNA yield, quality, and collection costs: comparison of methods for large-scale studies.
There is considerable interest in noninvasive and cost-effective methods for obtaining DNA in large-scale studies. In this randomized crossover study of 22 participants, we compared the DNA yield, quality, and associated costs of buccal cell DNA collected using cytobrushes (three brushes per collection) and swish (i.e., mouthwash) in self-administered procedures. There was a nonstatistically significant higher yield from the mouthwash compared with cytobrush collections (15.8 microg versus 12.0 microg, respectively; P = 0.53). PCR reactions that required short (0.3 kb) or intermediate (1.1 kb) DNA fragments were 100% successful for DNA from brush and mouthwash, whereas PCRs for reactions that required long fragments (7.8 kb) failed for all of the participants from cytobrush DNA and were 81% successful for DNA from the mouthwash source. The brush collections provided sufficient DNA for an estimated 150-225 PCR reactions requiring short and intermediate DNA fragments. The estimated per person costs for buccal brush DNA collections in large studies were less then half (8.50 dollars) those for the mouthwash method (18 dollars). In addition, we tested whether cytobrush instructions to rub cheeks before collection or collect cells only in the morning increased DNA yield and whether repeat brushings of the same cheek reduced DNA yield. These variations resulted in no significant differences in DNA yields. We conclude that the collection of DNA with cytobrushes using simple instructions is cost effective in large-scale studies, and yields sufficient quantity and quality of DNA for genotyping.
King
Irena B
IB
Cancer Prevention Research Program, Fred Hutchinson Cancer Research Center, Seattle, Washington, 98109-1024, USA.
Satia-Abouta
Jessie
J
Thornquist
Mark D
MD
Bigler
Jeannette
J
Patterson
Ruth E
RE
Kristal
Alan R
AR
Shattuck
Ann L
AL
Potter
John D
JD
White
Emily
E
Abouta
Jessie Satia
JS
D003362:Cost-Benefit Analysis; D018592:Cross-Over Studies; D004247:DNA / Q000302:isolation & purification*; D016172:DNA Fingerprinting; D004867:Equipment Design; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009061:Mouth Mucosa / Q000166:cytology*; D009067:Mouthwashes; D016133:Polymerase Chain Reaction / Q000191:economics; D011786:Quality Control; D013048:Specimen Handling / Q000191:economics
D009067:Mouthwashes; D004247:DNA