Collection of Genomic DNA from Adults in Epidemiological Studies by Buccal Cytobrush and Mouthwash

Author(s): Garcia-Closas M, Egan K, Abruzzo J, Newcomb P, Titus-Ernstoff L, Franklin T, Bender P, Beck J, Le Marchand L, Lum A, Alavanja M, Hayes RB, Rutter J, Buetow KH, Brinton L, Rothman N

Publication: Cancer Epidemiol Biomarkers Prev, 2001, Vol. 10, Page 687

PubMed ID: 11401920 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   To compare cytobrush samples sent by mail at room temperature without a transport medium with alcohol-based mouthwash samples sent by mail at room temperature for self-collection of buccal cells in terms of total human DNA yield, DNA integrity, and PCR success in amplifying beta-actin gene fragments.

   Summary of Findings:

   Total and human DNA yields from cytobrush samples were substantially lower than yields from mouthwash samples. In addition, cytobrush and mouthwash samples contain a mixture of DNA of human and nonhuman origin, and the amount of human DNA was higher in the mouthwash samples.

   Biospecimens

   • Cell - Buccal
   • Bodily Fluid - Saliva

   Preservative Types

   • Other Preservative

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   DNA	Electrophoresis
   DNA	PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Acquisition	Method of fluid acquisition	Cytological brush Swishing
   Biospecimen Acquisition	Method of cell acquisition	Cytological brush Swishing

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2. Study Purpose

   To compare phenol-chloroform DNA extraction to QIAamp DNA Blood Mini Kit, Puregene DNA Isolation Kit, and NaOH extraction in terms of total human DNA yield, DNA integrity, and PCR success in amplifying beta-actin gene fragments.

   Summary of Findings:

   Total and human DNA yields from cytobrush or mouthwash samples extracted by the QIAamp blood kit (Qiagen Inc.) were lower than the yields obtained by a phenol-chloroform extraction method. In addition, QIAamp DNA extracts were of lower molecular weight than phenol-chloroform extracts. NaOH-extracted DNA was of low molecular weight and presented difficulties for PCR amplification of beta-globin gene fragments. Total and human DNA yields as well as DNA integrity, PCR success rates, and number of attempts for a successful amplification were very similar for mouthwash samples extracted by Puregene or by phenol-chloroform.

   Biospecimens

   • Cell - Buccal
   • Bodily Fluid - Saliva

   Preservative Types

   • Other Preservative

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   DNA	DNA sequencing
   DNA	PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Analyte isolation method	Phenol-chloroform QIAamp DNA Blood Mini Kit Puregene DNA Isolation Kit NaOH extraction

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