Whole genome microarray analysis, from neonatal blood cards
Whole genome microarray analysis, from neonatal blood cards
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DOI：
10.1186/1471-2156-10-38
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发表时间：
2009-07-22
期刊：
影响因子：
2.9
通讯作者：
Shaw, Gary M.
中科院分区：
文献类型：
--
作者：
Hardin, Jill;Finnell, Richard H.;Shaw, Gary M.
背景：50年来，新生儿血液一直是出生缺陷筛查的标准，新生儿血液是从足跟贴上获得的，并干燥储存在纸卡片上。这种干燥的血液样品主要用于分析小分子分析物。最近，这种干血卡的DNA补体已用于靶向基因检测，例如囊性纤维化中的单核苷酸多态性。扩大这种测试，包括多基因性状，也许全基因组扫描，已被讨论作为一个正式的可能性。然而，到目前为止，可能从这样的干血卡获得的DNA的量一直是有限的，由于效率低下的DNA recovery technology.Results：采用一种新的技术，从一个标准的新生儿血卡有效的DNA释放。使用收集后平均储存十年的标准古特里卡，处理约1/40（th）的空气干燥的新生儿血液样本（两个3 mm穿孔）以获得质量和质量足以直接用于基于微阵列的全基因组扫描的DNA。使用该相同的DNA释放技术，还显示约1/250（th）的原始纯化的DNA（约1 ng）可以进行全基因组扩增，从而产生额外的微克扩增的DNA产物。扩增的DNA产物，然后用于微阵列分析，并产生了99%或更大的统计一致性的主要，未扩增的DNA samples.Conclusion：总之，这些数据表明，从不到10%的标准新生儿血液标本，干燥储存数年的古特里卡，可以支持一个程序的全基因组新生儿基因检测。
Background: Neonatal blood, obtained from a heel stick and stored dry on paper cards, has been the standard for birth defects screening for 50 years. Such dried blood samples are used, primarily, for analysis of small-molecule analytes. More recently, the DNA complement of such dried blood cards has been used for targeted genetic testing, such as for single nucleotide polymorphism in cystic fibrosis. Expansion of such testing to include polygenic traits, and perhaps whole genome scanning, has been discussed as a formal possibility. However, until now the amount of DNA that might be obtained from such dried blood cards has been limiting, due to inefficient DNA recovery technology.Results: A new technology is employed for efficient DNA release from a standard neonatal blood card. Using standard Guthrie cards, stored an average of ten years post-collection, about 1/40(th) of the air-dried neonatal blood specimen (two 3 mm punches) was processed to obtain DNA that was sufficient in mass and quality for direct use in microarray-based whole genome scanning. Using that same DNA release technology, it is also shown that approximately 1/250(th) of the original purified DNA (about 1 ng) could be subjected to whole genome amplification, thus yielding an additional microgram of amplified DNA product. That amplified DNA product was then used in microarray analysis and yielded statistical concordance of 99% or greater to the primary, unamplified DNA sample.Conclusion: Together, these data suggest that DNA obtained from less than 10% of a standard neonatal blood specimen, stored dry for several years on a Guthrie card, can support a program of genome-wide neonatal genetic testing.