Hollegaard Mads Vilhelm, Thorsen Poul, Norgaard-Pedersen Bent, Hougaard David Michael
Section of Neonatal Screening and Hormones, Statens Serum Institute, Copenhagen, Denmark.
Electrophoresis. 2009 Jul;30(14):2532-5. doi: 10.1002/elps.200800655.
Stored surplus of dried blood spot (DBS) samples from neonatal screening programs constitute a vast potential for large genetic epidemiological studies. However, age of the samples and the small amounts of DNA available may limit their usage. In this study we validate genotyping accuracy and efficiency of whole-genome-amplified DNA (wgaDNA) obtained from stored DBS samples, with reference to fresh genomic DNA from the same individuals. DBS samples from 29 volunteers, stored for up to 25 years, in the Danish Neonatal Screening Biobank were included and three DNA extraction methods, each using one 3.2 mm disk, were evaluated. Four whole-genome amplification kits, and one re-amplification kit, were used. Thirty-one SNPs were genotyped using the Sequenom platform and the wgaDNA samples calls were compared with their references for accuracy and efficiency evaluation. The genotype calls done blinded by the user had in many setups a 100% call- and concordance rate. Our results showed that genotyping performance is dependent on the combination of extraction procedure and amplification method, whereas years of storage did not seem to influence in this study. Based on these results we conclude that DBS samples should be considered a reliable and potential resource for future genotyping studies.
新生儿筛查项目中储存的干血斑（DBS）样本剩余量，为大规模基因流行病学研究提供了巨大潜力。然而，样本的保存时间以及可获取的DNA量较少，可能会限制其用途。在本研究中，我们参照同一研究对象的新鲜基因组DNA，验证了从储存的DBS样本中获取的全基因组扩增DNA（wgaDNA）的基因分型准确性和效率。我们纳入了丹麦新生儿筛查生物样本库中保存长达25年的29名志愿者的DBS样本，并评估了三种DNA提取方法，每种方法均使用一个3.2毫米滤纸片。我们使用了四种全基因组扩增试剂盒和一种再扩增试剂盒。利用Sequenom平台对31个单核苷酸多态性（SNP）进行基因分型，并将wgaDNA样本的分型结果与其对照进行比较，以评估准确性和效率。在许多实验设置中，由使用者盲法进行的基因型分型的检出率和一致性率均为100%。我们的结果表明，基因分型性能取决于提取程序和扩增方法的组合，而在本研究中，储存年限似乎并未产生影响。基于这些结果，我们得出结论，DBS样本应被视为未来基因分型研究的可靠且有潜力的资源。
