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首页> 外文期刊>BMC Medical Genomics >Saliva samples are a viable alternative to blood samples as a source of DNA for high throughput genotyping
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Saliva samples are a viable alternative to blood samples as a source of DNA for high throughput genotyping

机译：唾液样品是血液样品的可行替代品，可作为高通量基因分型的DNA来源

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Background The increasing trend for incorporation of biological sample collection within clinical trials requires sample collection procedures which are convenient and acceptable for both patients and clinicians. This study investigated the feasibility of using saliva-extracted DNA in comparison to blood-derived DNA, across two genotyping platforms: Applied Biosystems TaqmanTM and Illumina BeadchipTM genome-wide arrays. Method Patients were recruited from the Pharmacogenetics of Breast Cancer Chemotherapy (PGSNPS) study. Paired blood and saliva samples were collected from 79 study participants. The Oragene DNA Self-Collection kit (DNAgenotek?) was used to collect and extract DNA from saliva. DNA from EDTA blood samples (median volume 8 ml) was extracted by Gen-Probe, Livingstone, UK. DNA yields, standard measures of DNA quality, genotype call rates and genotype concordance between paired, duplicated samples were assessed. Results Total DNA yields were lower from saliva (mean 24 μg, range 0.2–52 μg) than from blood (mean 210 μg, range 58–577 μg) and a 2-fold difference remained after adjusting for the volume of biological material collected. Protein contamination and DNA fragmentation measures were greater in saliva DNA. 78/79 saliva samples yielded sufficient DNA for use on Illumina Beadchip arrays and using Taqman assays. Four samples were randomly selected for genotyping in duplicate on the Illumina Beadchip arrays. All samples were genotyped using Taqman assays. DNA quality, as assessed by genotype call rates and genotype concordance between matched pairs of DNA was high (>97%) for each measure in both blood and saliva-derived DNA. Conclusion We conclude that DNA from saliva and blood samples is comparable when genotyping using either Taqman assays or genome-wide chip arrays. Saliva sampling has the potential to increase participant recruitment within clinical trials, as well as reducing the resources and organisation required for multicentre sample collection.
机译：背景技术将生物样品收集纳入临床试验的增长趋势要求对患者和临床医生都方便且可接受的样品收集程序。这项研究调查了在两个基因分型平台上使用唾液提取的DNA与血液来源的DNA进行比较的可行性：Applied Biosystems Taqman TM 和Illumina Beadchip TM 全基因组阵列。方法从乳腺癌化疗药物遗传学（PGSNPS）研究中招募患者。从79名研究参与者中收集了成对的血液和唾液样本。 Oragene DNA自收集试剂盒（DNAgenotek？）用于从唾液中收集和提取DNA。来自英国利文斯通的Gen-Probe提取了EDTA血液样本（中位数8毫升）中的DNA。评估了成对重复样品之间的DNA产量，DNA质量的标准量度，基因型检出率和基因型一致性。结果唾液的总DNA产量（平均24μg，范围0.2-52μg）要比血液（平均210μg，范围58-577μg）低，并且在调整了所收集的生物材料的体积之后，仍然有2倍的差异。唾液DNA中的蛋白质污染和DNA片段化措施更大。 78/79唾液样品产生了足够的DNA，可用于Illumina Beadchip阵列和Taqman分析。在Illumina Beadchip阵列上随机选择四个样本进行基因分型。使用Taqman测定法对所有样品进行基因分型。通过基因型检出率和匹配的DNA对之间的基因型一致性来评估的DNA质量在血液和唾液来源的DNA中均达到较高水平（> 97％）。结论我们得出的结论是，使用Taqman分析或全基因组芯片阵列进行基因分型时，唾液和血液样本中的DNA具有可比性。唾液采样有可能增加临床试验中的参与者招募，并减少多中心样本收集所需的资源和组织。

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