Buccal cell DNA yield, quality, and collection costs: comparison of methods for large-scale studies.

Author(s): King IB, Satia-Abouta J, Thornquist MD, Bigler J, Patterson RE, Kristal AR, Shattuck AL, Potter JD, White E

Publication: Cancer Epidemiol Biomarkers Prev, 2002, Vol. 11, Page 1130-3

PubMed ID: 12376522 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine if DNA yield from cytobrush collection of buccal cells is affected by sequential collection, rubbing the cheek against the teeth or collection time of day. All specimens in this study were self-collected, transported by mail and stored at -80 degrees C until analysis.

   Summary of Findings:

   The yield of DNA collected by self-administered cytobrush was not significantly affected by prior collection from the same cheek. Further, yield was not increased by rubbing the cheek against the teeth prior to collection. No effect of collecting prior to the first meal of the day versus in the afternoon was observed. In conclusion, buccal cell DNA yield from cytobrush specimens was not significantly affected by the collection procedure used.

   Biospecimens

   • Cell - Buccal

   Preservative Types

   • Frozen

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   DNA	Spectrophotometry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Acquisition	Time of biospecimen collection	Before first meal of the day Anytime during the day
   Biospecimen Aliquots and Components	Aliquot sequential collection	1st collection 2nd collection
   Biospecimen Acquisition	Method of cell acquisition	Cytological brush Cheek rubbing prior to collection

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2. Study Purpose

   The purpose of this study was to compare DNA yield and quality from buccal cell specimens collected by alcohol based mouthwash or cytobrush methods. All collections were self-administered and transported by mail prior to storage at -80 degrees C. DNA quantification was done by spectrophotometer.

   Summary of Findings:

   Specimens collected by mouthwash had 30% more DNA then those collected by cytobrush, but this increase was not statistically significant (p=0.53). Furthermore, the variability between specimens was high. The A260/280 ratio for both methods ranged from 1.6-2.0. In all specimens collected, regardless of the method used, a 295 bp region of EHX and a 1.1 kB region of NAT2 were able to be amplified. Amplification of a 7.8 kB region of cyp2A6 was not possible from the cytobrush collected specimens, but occurred 81% of the time in specimens collected by mouthwash.

   Biospecimens

   • Cell - Buccal
   • Bodily Fluid - Saliva

   Preservative Types

   • Frozen

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   DNA	Spectrophotometry
   DNA	PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Acquisition	Method of fluid acquisition	Swishing Cytological brush
   PCR Specific	Length of gene fragment	295 bp 1.1 kb 7.8 kb
   Biospecimen Acquisition	Method of cell acquisition	Cytological brush Swishing

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