Institute of Molecular Biomedicine, Comenius University, Bratislava, Slovakia.
Clin Chem Lab Med. 2012 Apr;50(4):643-7. doi: 10.1515/CCLM.2011.814.
The non-invasive, flexible and easy sample collection makes saliva an interesting source of DNA for research and diagnostic purposes. The aim of our study was to find the most suitable collection method for biological material from the oral cavity and the most effective DNA isolation technique for further analytic applications.
DNA was isolated from swabs, Salivette saliva, whole saliva and samples collected with a commercial set for scraping of buccal cells. Phenol-chloroform extraction and isolation using a silica membrane based commercial kit were compared. Quantity of bacterial and human genomic DNA was estimated using real time PCR. The effects of storage conditions on DNA recovery were assessed.
Sample collection techniques significantly affected the quantity of DNA for both, silica membrane based and phenol-chloroform isolations. Whole saliva provided the largest number of bacterial and human genome copies after both extraction methods. Storage for 36 months at –20°C reduced recovery of human genomic DNA five times after silica membrane based extraction and 10 times after phenol-chloroform isolation.
Whole saliva was found to be the most suitable material for human and bacterial DNA isolation. Both compared methods are useful considering the quantity of extracted DNA.
唾液具有非侵入性、灵活性和易于采集样本的特点，因此成为研究和诊断目的的一种有趣的 DNA 来源。我们的研究目的是找到最适合从口腔采集生物材料的方法，以及最适合进一步分析应用的有效 DNA 分离技术。
从拭子、Salivette 唾液、全唾液以及使用商业刮取口腔细胞套件采集的样本中分离 DNA。比较了苯酚-氯仿抽提法和基于硅胶膜的商业试剂盒分离法。使用实时 PCR 估计细菌和人类基因组 DNA 的数量。评估了储存条件对 DNA 回收的影响。
两种提取方法的样本采集技术均显著影响 DNA 的数量。全唾液在两种提取方法后均提供了最多的细菌和人类基因组拷贝。在-20°C 下储存 36 个月后，硅胶膜提取后人类基因组 DNA 的回收减少了五倍，苯酚-氯仿分离后减少了十倍。
全唾液被发现是人类和细菌 DNA 分离的最适合材料。考虑到提取的 DNA 数量，两种比较方法都很有用。
