New saliva DNA collection method compared to buccal cell collection techniques for epidemiological studies
New saliva DNA collection method compared to buccal cell collection techniques for epidemiological studies
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DOI：
10.1002/ajhb.20586
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发表时间：
2007-05-01
影响因子：
2.9
通讯作者：
Demerath, Ellen W.
中科院分区：
文献类型：
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作者：
Rogers, Nikki L.;Cole, Shelley A.;Demerath, Ellen W.
流行病学研究可能需要非侵入性的非现场DNA收集方法。我们比较了使用全唾液采集设备(Oragene(商标)DNA采集试剂盒)和三种已建立的非侵入性方法(细胞刷子、泡沫拭子和口腔冲洗)获得的DNA产量和质量。每种方法都在我们中心的17名成年志愿者身上进行了测试，采用随机交叉收集设计，并使用重复测量统计进行分析。通过凝胶电泳法、分光光度法和聚合酶链式反应(PCR)扩增率评价DNA产量和质量。全唾液法的DNA得率(平均+/-SD=154.9+/-103.05 mU g，中位数=181.88)显著高于其他方法(口腔冲洗=54.74+/-41.72mU g，36.56；拭子=11.44+/-7.39mU g，10.72；细胞刷法12.66+/-6.19，13.22mU g)(均为P<；0.05)。口腔含漱液和全唾液样本提供的DNA质量最好，而细胞刷子和拭子样本提供的DNA质量较差，OD260/OD280和OD260/OD230比值较低。我们的结论是，与通常使用的口腔拭子和刷子技术相比，10毫升口腔含漱液样本和2毫升全唾液样本都能为遗传流行病学研究提供足够的DNA数量和更高质量的DNA。
Epidemiological studies may require noninvasive methods for off-site DNA collection. We compared the DNA yield and quality obtained using a whole-saliva collection device (Oragene (TM) DNA collection kit) to those from three established noninvasive methods (cytobrush, foam swab, and oral rinse). Each method was tested on 17 adult volunteers from our center, using a random crossover collection design and analyzed using repeated-measures statistics. DNA yield and quality were assessed via gel electrophoresis, spectophotometry, and polymerase chain reaction (PCR) amplification rate. The whole-saliva method provided a significantly greater DNA yield (mean +/- SD = 154.9 +/- 103.05 mu g, median = 181.88) than the other methods (oral rinse = 54.74 +/- 41.72 mu g, 36.56; swab = 11.44 +/- 7.39 mu g, 10.72; cytobrush 12.66 +/- 6.19, 13.22 mu g) (all pairwise P < 0.05). Oral-rinse and whole-saliva samples provided the best DNA quality, whereas cytobrush and swab samples provided poorer quality DNA, as shown by lower OD260/OD280 and OD260/OD230 ratios. We conclude that both a 10-ml oral-rinse sample and 2-ml whole-saliva sample provide sufficient DNA quantity and better quality DNA for genetic epidemiological studies than do the commonly used buccal swab and brush techniques.