Evaluation of Mailed Pediatric Buccal Cytobrushes for Use in a Case-Control Study of Birth Defects
Evaluation of Mailed Pediatric Buccal Cytobrushes for Use in a Case-Control Study of Birth Defects
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DOI：
10.1002/bdra.20829
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发表时间：
2011-07-01
影响因子：
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通讯作者：
Rasmussen, Sonja A.
中科院分区：
文献类型：
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作者：
Gallagher, Margaret L.;Sturchio, Cynthia;Rasmussen, Sonja A.
背景技术背景：颊细胞收集是一种方便的DNA收集方法，然而，很少有人注意到从儿科人群中获得的DNA的质量。本研究的目的是确定一种改良的细胞刷收集方法对婴儿口腔DNA的产量和质量的影响，作为出生缺陷的基于人群的病例对照研究的一部分。方法：从1997年至2003年使用标准收集方法从婴儿、母亲和父亲收集细胞刷，并从2003年至今使用允许在收集后风干细胞刷的修改方案。分别采用定量PCR和短串联重复序列（STR）基因分型方法对1057个细胞刷的DNA产量和质量进行了评估。研究结果：来自婴儿的风干细胞刷的中位DNA产量（1300 ng）和STR完成率（99.5%）高于标准收集方法细胞刷（分别为60 ng和59.5%）。一个子集的DNA等分试样进行基因分型的六个单核苷酸多态性（SNP）。来自两种收集方法的通过质量方案（DNA浓度>1 ng/μ l，并且成功扩增>= 1个STR）的等分试样具有高基因型完成率（99-100%）。全基因组扩增后，风干标本的中位DNA产量比标准标本高两倍以上（p < 0.001）。结论：从婴儿收集的口腔DNA的产量和质量通过使用结合空气干燥的方法得到改善;然而，如果制定严格的质量控制程序，则通过这两种方法收集的DNA适合用于基因分型。这些发现可能有助于未来出生缺陷和其他不良儿科结局的流行病学研究。出生缺陷研究（A部分）91：642-648，2011年。(C)2011 Wiley-Liss，Inc.
BACKGROUND: Buccal cell collection is a convenient DNA collection method; however, little attention has been given to the quality of DNA obtained from pediatric populations. The purpose of this study was to determine the effect of a modified cytobrush collection method on the yield and quality of infant buccal DNA collected as part of a population-based case-control study of birth defects. METHODS: Cytobrushes were collected from infants, mothers, and fathers using a standard collection method in 1997 to 2003 and a modified protocol that allows air-drying of the cytobrushes after collection from 2003 to the present. Yield and quality of DNA from 1057 cytobrushes was assessed by quantitative PCR and short tandem repeat (STR) genotyping, respectively. RESULTS: Air-dried cytobrushes from infants had higher median DNA yields (1300 ng) and STR completion rates (99.5%) than standard collection method cytobrushes (60 ng and 59.5%, respectively). A subset of DNA aliquots was genotyped for six single nucleotide polymorphisms (SNPs). Aliquots from both collection methods that passed the quality protocol (DNA concentration >1 ng/mu l, and successful amplification of >= 1 STR) had high genotype completion rates (99-100%). The median DNA yield following whole genome amplification was more than twofold higher for air-dried than standard collection specimens (p < 0.001). CONCLUSION: Yield and quality of buccal DNA collected from infants are improved by using a method that incorporates air-drying; however, DNA collected by both methods is suitable for genotyping if stringent quality control procedures are instituted. These findings may be helpful for future epidemiologic studies of birth defects and other adverse pediatric outcomes. Birth Defects Research (Part A) 91:642-648, 2011. (C) 2011 Wiley-Liss, Inc.