【摘要】 目的 :寻求一种能方便、简捷、有效地获取患者DNA的方法 ,以用于检测患者基因多态性。方法 :采用 4种方法获取患者DNA ,即静脉血中用酚 -氯仿法抽提 ,指尖血血痕中用Chelex 10 0法抽提 ,颊粘膜拭子中用Chelex 10 0法抽提及直接用干燥血痕作为PCR扩增的DNA模板。对同一个体分别用 4种来源的DNA检测其IL 1基因多态性 ,比较结果的特异性和敏感性。结果 :颊粘膜拭子抽提DNA作基因多态性分析特异性高、敏感性强 ,优于静脉血酚 -氯仿法抽提DNA ,且取颊粘膜拭子无创、方便。Chelex 10 0法抽提DNA比酚 -氯仿法更简单、快速。结论 :颊粘膜拭子结合Chelex 10 0法抽提DNA适于对牙周炎患者作基因多态性分析更多还原

【Abstract】 Objective: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. Methods: 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. Results: 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 63.8μg±18.7μg, which is enough for 10 PCR reactions. Conclusion: Buccal swab appears to be an excellent source of DNA for detection of polymorphisms of human IL 1 gene.\;更多还原