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首页> 外文期刊>Psychiatric genetics >Development and validation of the high-quality 'rapid method for swab' to genotype the HTTLPR serotonin transporter (SLC6A4) promoter polymorphism.
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Development and validation of the high-quality 'rapid method for swab' to genotype the HTTLPR serotonin transporter (SLC6A4) promoter polymorphism.

机译：高质量“拭子快速方法”的开发和验证，用于对HTTLPR血清素转运蛋白（SLC6A4）启动子多态性进行基因分型。

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BACKGROUND: The importance of genetic variation to the etiology of neuropsychiatric disorders is well established and is currently being examined for diagnosis and treatment. The most popular method of obtaining material for genotype analysis, high-yielding DNA extraction from blood, has several limitations, including invasiveness, need for skilled individuals to collect material, and requirement for cold storage. Saliva sampling is noninvasive and trained personnel are less necessary, but it still requires a relatively high level of subject compliance. Buccal mucosa cells sampling is almost completely noninvasive, reducing compliance issues significantly. Samples collected have been shown to produce usable DNA after shipment through conventional mail. The DNA produced by rapid elution of these swabs in chaotropic buffers is, however, of limited quality and low purity. OBJECTIVE: Our aim was to develop a rapid, economical, and environmentally safe method for extraction of high-quality genomic DNA, which can be used to determine clinically important genotypes from trace quantity samples and which has sufficient yield for multiple assays. METHODS: We developed a method of extracting high-quality genomic DNA from buccal swab, which we termed the 'rapid method for swab' (RMS). We compared RMS with two established procedures, specifically the original rapid method and the commercially available Buccal Amp method. We assessed the generated genomic DNAs by their (i) quality, (ii) quantity, (iii) restriction enzyme digestibility, and (iv) PCR-based genotyping in addition to time, cost, and environmental impact of the procedures. MAIN RESULTS: DNA generated by RMS was of higher purity than that by Buccal Amp. RMS is nonenzymatic and does not use strong chaotropic salts or extreme pH. We also showed the suitability of RMS-DNA for LA/LG genotyping as generated by PCR using 7-deaza-dGTP. CONCLUSION: The RMS procedure is novel, efficient, safe, and yields sufficient material for multiple genotyping analyses. The RMS produces DNA of high quality from a single human buccal swab. RMS is a noninvasive technique and particularly suitable for children and older individuals and in field collection settings.
机译：背景：遗传变异对神经精神疾病的病因学的重要性已得到充分确立，目前正在对其进行诊断和治疗。从基因提取分析中获取材料的最流行方法是从血液中提取高产DNA，但存在一些局限性，包括侵入性，需要熟练的技术人员来收集材料以及需要冷藏。唾液采样是非侵入性的，培训的人员较少，但是仍然需要相对较高的受试者依从性。颊粘膜细胞采样几乎完全是非侵入性的，从而大大减少了依从性问题。通过常规邮件运输后，收集到的样品已显示出可产生有用的DNA。但是，通过在离液缓冲液中快速洗脱这些拭子产生的DNA质量有限且纯度低。目的：我们的目的是开发一种快速，经济，环境安全的高质量基因组DNA提取方法，该方法可用于从痕量样品中确定临床上重要的基因型，并具有足够的产率进行多种测定。方法：我们开发了一种从颊拭子中提取高质量基因组DNA的方法，我们称之为“拭子快速方法”（RMS）。我们将RMS与两个既定程序进行了比较，特别是原始的快速方法和市售的Buccal Amp方法。我们评估了产生的基因组DNA的质量，过程，时间，成本和环境影响，这些因素包括：（i）质量，（ii）数量，（iii）限制酶消化率和（iv）基于PCR的基因型。主要结果：RMS产生的DNA纯度高于Buccal Amp。 RMS是非酶促的，不使用强离液盐或极端pH。我们还显示了使用7-deaza-dGTP通过PCR生成的RMS-DNA对LA / LG基因分型的适用性。结论：RMS程序新颖，高效，安全，可为多种基因分型分析提供足够的材料。 RMS可从单个人的口腔拭子中产生高质量的DNA。 RMS是一种非侵入性技术，特别适合儿童和年龄较大的人以及野外采集场所。

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