Morphology Department, Institute of Biology, Universidade Federal de Pelotas, Pelotas, Brazil.
BMC Med Res Methodol. 2012 May 4;12:65. doi: 10.1186/1471-2288-12-65.
Large epidemiological studies in DNA biobanks have increasingly used less invasive methods for obtaining DNA samples, such as saliva collection. Although lower amounts of DNA are obtained as compared with blood collection, this method has been widely used because of its more simple logistics and increased response rate. The present study aimed to verify whether a storage time of 8 months decreases the quality of DNA from collected samples.
Saliva samples were collected with an OrageneTM DNA Self-Collection Kit from 4,110 subjects aged 14-15 years. The samples were processed in two aliquots with an 8-month interval between them. Quantitative and qualitative evaluations were carried out in 20% of the samples by spectrophotometry and genotyping. Descriptive analyses and paired t-tests were performed.
The mean volume of saliva collected was 2.2 mL per subject, yielding on average 184.8 μg DNA per kit. Most samples showed a Ratio of OD differences (RAT) between 1.6 and 1.8 in the qualitative evaluation. The evaluation of DNA quality by TaqMan®, High Resolution Melting (HRM), and restriction fragment length polymorphism-PCR (RFLP-PCR) showed a rate of success of up to 98% of the samples. The sample store time did not reduce either the quantity or quality of DNA extracted with the Oragene kit.
The study results showed that a storage period of 8 months at room temperature did not reduce the quality of the DNA obtained. In addition, the use of the Oragene kit during fieldwork in large population-based studies allows for DNA of high quantity and high quality.
在 DNA 生物库的大型流行病学研究中，越来越多地采用了获取 DNA 样本的非侵入性方法，例如唾液采集。虽然与血液采集相比，获得的 DNA 量较少，但由于其更简单的物流和更高的响应率，这种方法已被广泛使用。本研究旨在验证储存 8 个月是否会降低采集样本的 DNA 质量。
从 4110 名 14-15 岁的受试者中采集唾液样本，使用 OrageneTM DNA 自采集试剂盒。这些样本在间隔 8 个月的时间内分成两份进行处理。通过分光光度法和基因分型对 20%的样本进行定量和定性评估。进行描述性分析和配对 t 检验。
每位受试者采集的唾液平均体积为 2.2 毫升，每个试剂盒平均可获得 184.8 微克 DNA。大多数样本在定性评估中 RAT 值在 1.6 到 1.8 之间。通过 TaqMan®、高分辨率熔解（HRM）和限制性片段长度多态性-PCR（RFLP-PCR）评估 DNA 质量，样本的成功率高达 98%。样本储存时间并未降低 Oragene 试剂盒提取的 DNA 的数量或质量。
研究结果表明，在室温下储存 8 个月不会降低所获得的 DNA 的质量。此外，在基于人群的大型研究中使用 Oragene 试剂盒可以获得高质量、高数量的 DNA。
