评估用于遗传易感性研究的颊细胞采集方案。

Evaluation of buccal cell collection protocols for genetic susceptibility studies.

机构信息

University of California at Berkeley, School of Public Health, 140 Warren Hall, Berkeley CA 94720-7360, USA.

出版信息

Biomarkers. 2001;6(6):448-54. doi: 10.1080/13547500110057416.

Abstract

Buccal cells are increasingly used as a source of quality DNA to improve participation rates in molecular studies. Here, three buccal cell collection protocols were compared to determine factors affecting the yield of cells, total DNA per sample, and DNA yield per cell. In addition, kinetic quantitative polymerase chain reaction (PCR) (TaqMan™) was used to quantify human DNA available for PCR. The method of collection used influenced the overall DNA yield per sample. The collection buffer used influenced the number of cells but not the overall DNA yield per sample. Repeated freezing and thawing did not affect overall DNA yield per sample, DNA yield per cell, or the total number of cells collected. Mouthwashes had the highest DNA yield per sample (20.8 μg) compared with cytobrush samples (1.9 μg from three cytobrushes) and tongue depressors (0.8 μg from three tongue depressors). However, mouthwash samples may contain significant non-human DNA and other contaminants that could interfere with some molecular studies. Spectrometry grossly overestimated the total DNA recovered from mouthwash samples compared with fluorometry or quantitative PCR.

摘要

颊细胞越来越多地被用作获取高质量 DNA 的来源，以提高分子研究的参与率。在这里，比较了三种颊细胞采集方案，以确定影响细胞产量、每个样本总 DNA 量和每个细胞 DNA 产量的因素。此外，还使用实时定量聚合酶链反应（PCR）（TaqMan™）来定量可用于 PCR 的人 DNA。所使用的采集方法会影响每个样本的总 DNA 产量。采集缓冲液会影响细胞数量，但不会影响每个样本的总 DNA 产量。反复冻融不会影响每个样本的总 DNA 产量、每个细胞的 DNA 产量或采集的细胞总数。与细胞刷样本（三个细胞刷的 1.9 μg）和压舌器样本（三个压舌器的 0.8 μg）相比，漱口剂的每个样本 DNA 产量最高（20.8 μg）。然而，漱口剂样本可能含有大量的非人类 DNA 和其他污染物，可能会干扰某些分子研究。与荧光计或定量 PCR 相比，光谱法严重高估了从漱口剂样本中回收的总 DNA。