Effects of source of DNA on genotyping success rates and allele percentages in the Preschoolers with Attention-Deficit/Hyperactivity Disorder Treatment Study (PATS).
Effects of source of DNA on genotyping success rates and allele percentages in the Preschoolers with Attention-Deficit/Hyperactivity Disorder Treatment Study (PATS).
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DOI：
10.1089/cap.2007.0076
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发表时间：
2007-11
影响因子：
1.9
通讯作者：
J. Swanson;R. Moyzis;J. McGough;J. McCracken;M. Riddle;S. Kollins;L. Greenhill;H. Abikoff;T. Wigal;S. Wigal;K. Posner;Anne M. Skrobala;M. Davies;J. Ghuman;C. Cunningham;B. Vitiello;A. Stehli;S. Smalley;D. Grady
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作者：
J. Swanson;R. Moyzis;J. McGough;J. McCracken;M. Riddle;S. Kollins;L. Greenhill;H. Abikoff;T. Wigal;S. Wigal;K. Posner;Anne M. Skrobala;M. Davies;J. Ghuman;C. Cunningham;B. Vitiello;A. Stehli;S. Smalley;D. Grady
目的在被诊断为注意缺陷多动障碍（ADHD）的儿童及其父母中，他们是学龄前ADHD治疗研究（PATS）的参与者，我们评估了DNA来源的影响（来自口腔或血细胞）对三个候选基因中五个多态性的基因分型成功率和等位基因百分比的影响（DAT 1、DRD 4和SNAP 25）在PATS药物遗传学研究中对刺激性药物的反应进行了研究。方法在基线评估时，241名个体（113名先证者和128名父母）同意参与; 144名个体（52名先证者和92名父母）提供静脉穿刺血样，97名个体（61名先证者和36名父母）提供颊拭子样本作为DNA分离标本。对DRD 4基因外显子III的48 bp VNTR、DAT 1基因3 '端非翻译区的40 bp VNTR、DRD 4基因启动子区的TDP 120 bp重复、SNAP-25基因TC-1069 SNP、和SNAP-25基因TG-1065 SNP。使用标准程序对这五种多态性中的每一种进行个体基因分型。结果使用2004年可用的方法，来自血细胞的DNA的基因分型成功率平均比来自口腔细胞的DNA的基因分型成功率高得多（例如，91% vs.先证者54%）。对于一些多态性（DRD 4-VNTR、DRD 4-TDP和SNAP 25-TC SNP），等位基因比例也因DNA的血液与口腔来源而变化（例如，DRD 4基因的7个重复等位基因分别为26.5%和18.6%）。结论：基于口腔DNA的基因分型成功率比血细胞低得多，这可能是由于这两种来源的DNA的质量。观察到的等位基因比例的来源差异可能是由于与样本收集方式（从颊拭子或静脉穿刺）的选择相关的自我选择，或基于DNA质量差异选择性检测某些等位基因。
OBJECTIVE In children diagnosed with attention-deficit/hyperactivity disorder (ADHD) and their parents, who were participants of the Preschool ADHD Treatment Study (PATS), we assessed the effect of source of DNA (from buccal or blood cells) on the genotyping success rate and allele percentages for the five polymorphisms in three candidate genes (DAT1, DRD4, and SNAP 25) investigated in the PATS pharmacogenetic study of response to stimulant medication. METHOD At baseline assessment, 241 individuals (113 probands and 128 parents) consented to participate; 144 individuals (52 probands and 92 parents) provided blood samples from venipuncture, and 97 individuals (61 probands and 36 parents) provided buccal samples from cheek swab as specimens for isolation of DNA. Three types of polymorphisms-variable number of tandem repeat (VNTR) polymorphism, tandem duplication polymorphism (TDP), and single nucleotide polymorphism (SNP)-were evaluated, including the DRD4 gene 48-bp VNTR in exon III, the DAT1 gene 40-bp VNTR in 3'-untranslated region, the DRD4 gene TDP 120-bp duplication in the promoter region, the SNAP-25 gene TC-1069 SNP, and the SNAP-25 gene TG-1065 SNP. Standard procedures were used to genotype individuals for each of these five polymorphisms. RESULTS Using the methods available in 2004, the genotyping success rate was on the average much greater for DNA from blood cells than buccal cells (e.g., 91% vs. 54% in probands). For some polymorphisms (DRD4-VNTR, DRD4-TDP, and SNAP25-TC SNP), allele proportion also varied by blood versus buccal source of DNA (e.g., 26.5% vs. 18.6% for the 7-repeat allele of the DRD4 gene). CONCLUSIONS The much lower success rate for genotyping based on DNA from buccal than blood cells is likely due to the quality of DNA derived from these two sources. The observed source differences in allele proportion may be due to self-selection related to choice of how specimens were collected (from cheek swab or venipuncture), or to a selective detection of some alleles based on differences in DNA quality.