Duan H, Zhang J, Huang P, Zhang Y
College of Stomatology, West China University of Medical Sciences.
Hua Xi Kou Qiang Yi Xue Za Zhi. 2001 Feb;19(1):11-3.
PCR-RFLP based techniques have become standard procedures for gene polymorphism screening. Peripheral venous blood is currently the most commonly employed source of DNA for human genome analysis. However, it has many practical disadvantage and inherent limitations to use blood as DNA source. Blood sampling is invasive, painful and involves a potential risk of contamination with hepatitis. The classic procedure to extract genomic DNA from whole blood is phenol/chloroform technique, which is relative complicated and time consuming. Therefore, this study was performed to try an alternative instead of using blood as DNA source for gene polymorphism analysis.
Four methods were employed to obtain DNA from the same subject including DNA from venous blood through phenol/chloroform extraction, DNA from dried blood spot through Chelex 100 technique, DNA from buccal swab through Chelex 100 technique and the dried blood spot directly as DNA template for PCR. Then, these various forms of DNA were used in PCR RFLP procedure to analyze IL-1 gene polymorphisms, and their specificity and sensitivity were evaluated.
Our results indicate that both the buccal swab and the blood based assays reached complete concordance in typing the IL-1 gene polymorphisms, while the Chelex 100 procedure for extracting DNA from buccal swab is much simpler and more rapid. It is non-invasive to get buccal swab. The amount of DNA obtained through one buccal swab is 63.8 micrograms +/- 18.7 micrograms, which is enough for 10 PCR reactions.
Buccal swab appears to be an excellent source of DNA for detection of polymorphisms of human IL-1 gene.
基于聚合酶链反应-限制性片段长度多态性（PCR-RFLP）的技术已成为基因多态性筛查的标准方法。外周静脉血是目前人类基因组分析中最常用的DNA来源。然而，将血液用作DNA来源存在许多实际缺点和固有局限性。采血具有侵入性、会带来疼痛，并且存在感染肝炎的潜在风险。从全血中提取基因组DNA的经典方法是酚/氯仿技术，该方法相对复杂且耗时。因此，本研究旨在尝试一种替代方法，不使用血液作为基因多态性分析的DNA来源。
采用四种方法从同一受试者获取DNA，包括通过酚/氯仿提取法从静脉血中获取DNA、通过Chelex 100技术从干血斑中获取DNA、通过Chelex 100技术从口腔拭子中获取DNA以及直接将干血斑作为PCR的DNA模板。然后，将这些不同形式的DNA用于PCR-RFLP程序以分析白细胞介素-1（IL-1）基因多态性，并评估其特异性和敏感性。
我们的结果表明，口腔拭子和基于血液的检测方法在对IL-1基因多态性进行分型时完全一致，而通过Chelex 100技术从口腔拭子中提取DNA的方法要简单得多且速度更快。获取口腔拭子是非侵入性的。通过一次口腔拭子获得的DNA量为63.8微克±18.7微克，足以进行10次PCR反应。
口腔拭子似乎是检测人类IL-1基因多态性的极佳DNA来源。
