Ng Daniel P K, Koh David, Choo Serena, Chia Kee-Seng
Department of Community, Occupational and Family Medicine, National University of Singapore, Singapore.
Clin Chim Acta. 2006 May;367(1-2):81-5. doi: 10.1016/j.cca.2005.11.024. Epub 2006 Jan 4.
Saliva is a potentially useful but untapped source of genomic DNA for genetic epidemiological studies. However, current commercial methods are mainly concerned with DNA extraction and do not address important issues concerning saliva preservation and storage. As such, we evaluated how various saliva storage conditions affected DNA yield and quality obtained using a new commercially available method that proposes to integrate these aspects in a single kit.
The conditions involved the extraction of the DNA immediately after saliva collection (condition 1) or when stored at air-conditioned room temperature (20 degrees C) for 1 month (condition 2) and 6 months (condition 3) as well as at -80 degrees C for 6 months (condition 4). The effect of incorporating an additional incubation of saliva samples at 30 degrees C for 2 weeks was also examined.
Overall average DNA yield from 2 ml of saliva was 35.5 microg (8.5-85.2 microg). DNA yield was unaffected by incubation of saliva at 30 degrees C but DNA yield under condition 3 was significantly higher compared to conditions 1 and 2. OD260/280 values were acceptable and comparable across all conditions. Differences in storage conditions did not impact DNA quality in real time PCR experiments and genotyping fidelity remained undiminished.
Saliva is a viable alternative source of human genomic DNA for genetic epidemiological studies and that this new commercial method and possibly other related techniques can be effective means towards this end.
唾液是遗传流行病学研究中一种潜在有用但尚未开发的基因组DNA来源。然而,目前的商业方法主要关注DNA提取,并未解决唾液保存和储存的重要问题。因此,我们评估了各种唾液储存条件对使用一种新的市售方法获得的DNA产量和质量的影响,该方法提议将这些方面整合在一个试剂盒中。
这些条件包括在唾液采集后立即提取DNA(条件1),或在空调室温(20摄氏度)下储存1个月(条件2)和6个月(条件3)以及在-80摄氏度下储存6个月(条件4)。还研究了在30摄氏度下对唾液样本额外孵育2周的影响。
2毫升唾液的总体平均DNA产量为35.5微克(8.5 - 85.2微克)。DNA产量不受唾液在30摄氏度下孵育的影响,但条件3下的DNA产量与条件1和2相比显著更高。所有条件下的OD260/280值均可接受且相当。储存条件的差异在实时PCR实验中不影响DNA质量,基因分型保真度也未降低。
唾液是遗传流行病学研究中人类基因组DNA的可行替代来源,这种新的商业方法以及可能的其他相关技术可以有效地实现这一目的。