不同方法提取石蜡包埋组织DNA的比较分析

A comparative analysis of different DNA extraction methods from paraffin-embedded tissues

目的:比较福尔马林固定石蜡包埋组织(FFPET)中3种提取DNA的方法对DNA质量和回收率的影响,探讨一种高效、简便、实用的石蜡组织中DNA提取方法。方法:选取新疆医科大学第一附属医院2004～2007年手术切除的10%甲醛固定石蜡包埋甲状腺癌组织标本,分别以试剂盒法、改良TES水浴脱蜡-酚氯仿法、改良TES水浴脱蜡-试剂盒法提取其DNA,然后进行电泳分析。结果:改良TES水浴脱蜡-酚氯仿提纯DNA法所得样本DNA的OD260/OD280比值为(1.82±0.15),改良TES水浴脱蜡-试剂盒提纯DNA法为(1.86±0.17),两者比较无明显差异(P>0.05);分别与试剂盒法(1.75±0.14)相比,差异有统计学意义(P<0.01)。改良TES水浴脱蜡-试剂盒提纯DNA法、改良TES水浴脱蜡-酚氯仿提纯DNA法所得DNA产量和质量均高于试剂盒法。结论:改良TES水浴脱蜡-试剂盒提纯法简单快捷,是较理想的石蜡包埋组织DNA提纯方法。

Objective： In order to identify the optimal method for DNA extraction from formalin-fixed, paraffin-embedded tissues, we compared three methods. Methods： The formalin-fixed, paraffin-embedded thyroid cancerous tissue specimen were selected in the First Affiliated Hospital of Xinjiang Medical University＇s operation specimen from 2004 to 2007, and divided into three groups according to the method for DNA extraction. Three methods included： （1） commercial genomic DNA extraction mini kit; （2） improved TES method of de-waxing with water-bath and conventional phenol - chloroform protocol; （3） im proved TES method of de-waxing with water-bath and commercial genomic DNA extraction mini kit. The quality of obtained DNA was analyzed by electrophoresis amplification. Results： For paraffin-embedded tis- sue specimens, OD260/OD280 ratio of DNA obtained by using improved TES method of de-waxing with water-bath and conventional phenol-chloroform protocol were （1.82±0.15） and （1.86±0.17） by using improved TES method of de-waxing with water-bath and commercial genomic DNA extraction mini kit, the result had no statically signification （P ±0.05）, but both have more statically signification （P 〈0.01）, compare with commercial genomic DNA extraction mini kit （1.75±0.14）. Both the improved TES method of de-waxing with water-bath and conventional phenol-chloroform protocol and improved TES method of de-waxing with water-bath and commercial genomic DNA extraction mini kit method were better methods for DNA isolation from formalin-fixed, paraffin-embedded tissue specimens, both the aspect of quantity and quality. Conclusion： Improved TES method of de-waxing with water-bath and commercial genomic DNA extraction mini kit is an optimal method for extraction of DNA from formalin-fixed, paraffin-embedded tissues.