Poynter Jenny N, Ross Julie A, Hooten Anthony J, Langer Erica, Blommer Crystal, Spector Logan G
BMC Genet. 2013 Aug 12;14:69. doi: 10.1186/1471-2156-14-69.
Collection of high-quality DNA is essential for molecular epidemiology studies. Methods have been evaluated for optimal DNA collection in studies of adults; however, DNA collection in young children poses additional challenges. Here, we have evaluated predictors of DNA quantity in buccal cells collected for population-based studies of infant leukemia (N = 489 mothers and 392 children) and hepatoblastoma (HB; N = 446 mothers and 412 children) conducted through the Children's Oncology Group. DNA samples were collected by mail using mouthwash (for mothers and some children) and buccal brush (for children) collection kits and quantified using quantitative real-time PCR. Multivariable linear regression models were used to identify predictors of DNA yield.
Median DNA yield was higher for mothers in both studies compared with their children (14 μg vs. <1 μg). Significant predictors of DNA yield in children included case-control status (β = -0.69, 50% reduction, P = 0.01 for case vs. control children), brush collection type, and season of sample collection. Demographic factors were not strong predictors of DNA yield in mothers or children in this analysis.
The association with seasonality suggests that conditions during transport may influence DNA yield. The low yields observed in most children in these studies highlight the importance of developing alternative methods for DNA collection in younger age groups.
高质量DNA的收集对于分子流行病学研究至关重要。在成人研究中,已对获取最佳DNA的方法进行了评估;然而,幼儿的DNA收集带来了额外的挑战。在此,我们评估了通过儿童肿瘤学组进行的婴儿白血病(N = 489名母亲和392名儿童)和肝母细胞瘤(HB;N = 446名母亲和412名儿童)基于人群研究中收集的颊细胞中DNA数量的预测因素。DNA样本通过邮寄方式收集,使用漱口水(用于母亲和部分儿童)和颊刷(用于儿童)收集试剂盒,并使用定量实时PCR进行定量。多变量线性回归模型用于确定DNA产量的预测因素。
在两项研究中,母亲的DNA产量中位数均高于其子女(14μg对<1μg)。儿童DNA产量的显著预测因素包括病例对照状态(β = -0.69,降低50%,病例组儿童与对照组儿童相比P = 0.01)、刷取收集类型和样本收集季节。在该分析中,人口统计学因素并非母亲或儿童DNA产量的有力预测因素。
与季节性的关联表明运输过程中的条件可能会影响DNA产量。在这些研究中,大多数儿童的产量较低,这凸显了为较年轻年龄组开发替代DNA收集方法的重要性。