DNA yield and quality of saliva samples and suitability for large-scale epidemiological studies in children
DNA yield and quality of saliva samples and suitability for large-scale epidemiological studies in children
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DOI：
10.1038/ijo.2011.43
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发表时间：
2011-04-01
影响因子：
4.9
通讯作者：
Pitsiladis, Y. P.
中科院分区：
文献类型：
--
作者：
Koni, A. C.;Scott, R. A.;Pitsiladis, Y. P.
目的：评价两种唾液DNA采集方法在一个大型、综合、多中心的欧洲儿童生物材料采集项目中的DNA产量和质量。设计：幼儿横断面多中心比较研究。方法：收集来自8个欧洲国家的14019名2-9岁儿童的唾液样本，这些儿童和婴儿的饮食和生活方式引起的健康影响的识别和预防（IDEFICS）研究。这包括从会吐痰的儿童身上收集2毫升唾液，或者用海绵从不能吐痰的幼儿的口腔内部收集整个唾液和口腔粘膜细胞。样本在每个参与中心集中组装，随后送往格拉斯哥大学进行DNA提取和生物银行。在基因分型之前，选择4678个样本（约占样本个体的33%）进行DNA提取。结果：全唾液采集法DNA得率高于海绵采集法（平均+/- s.d，唾液：20.95 +/- 2.35 mu g，海绵：9.13 +/- 2.25 mu g, P < 0.001）。用A(260)/A（280）测量的DNA质量在两种收集方法中是相似的。最小基因型呼叫成功率为95%，表明两种方法均可为TaqMan等位基因鉴别试验提供高质量的DNA。结论：我们的研究结果表明，与辅助海绵收集法相比，全唾液收集法的DNA产量更高。然而，这两种收集方法都为大规模遗传流行病学研究提供了足够数量和质量的DNA。国际肥胖杂志（2011）35,S113-S118；doi: 10.1038 / ijo.2011.43
Objective: To evaluate two saliva collection methods for DNA yield and quality as applied to a large, integrated, multicentre, European project involving the collection of biological material from children.Design: Cross-sectional multicentre comparative study in young children.Methods: Saliva samples were collected from 14 019 children aged 2-9 years from eight European countries participating in the IDEFICS (Identification and prevention of dietary-and lifestyle-induced health effects in children and infants) study. This involved either the collection of 2 ml of saliva from children who were able to spit, or using a sponge to collect whole saliva and buccal mucosal cells from the inside of the mouth of younger children unable to spit. Samples were assembled centrally in each participating centre and subsequently despatched for DNA extraction and biobanking to the University of Glasgow. A subgroup of 4678 samples (similar to 33% of sampled individuals) were chosen for DNA extraction before genotyping.Results: The whole-saliva collection method resulted in a higher DNA yield than the sponge collection method (mean +/- s.d.; saliva: 20.95 +/- 2.35 mu g, sponge: 9.13 +/- 2.25 mu g; P < 0.001). DNA quality as measured by A(260)/A(280) was similar for the two collection methods. A minimum genotype calling success rate of 95% showed that both methods provide good-quality DNA for genotyping using TaqMan allelic discrimination assays.Conclusions: Our results showed higher DNA yield from the whole-saliva collection method compared with the assisted sponge collection. However, both collection methods provided DNA of sufficient quantity and quality for large-scale genetic epidemiological studies. International Journal of Obesity (2011) 35, S113-S118; doi:10.1038/ijo.2011.43