CAJ | 학술논문

目的：依据血浆中存在游离DNA的理论，采用磁珠作为固相吸附载体并使用特定设计的试剂体系及提取流程，建立一种简便、高效提取尿液样本中游离甲基化DNA的方法，评价其作为用于尿液样本的甲基化基因检测技术的可行性。方法使用磁珠法提取40例成人尿液中游离甲基化DNA，进行甲基化修饰后，紫外分光光度计测定DNA的浓度和纯度。结果结果显示，提取50 mL尿液可得61~200 ng/μL的甲基化DNA，OD260/280为1.8±0.05。使用甲基化阳性对照DNA引物进行PCR及电泳后可见目的条带，说明其纯度可以满足后续甲基化基因检测和PCR等操作要求。结论尿液中确实存在游离甲基化DNA，同时磁珠法提取尿液游离DNA提取过程简单，且提取纯度高，是一种高效的提取方法。
목적：의거혈장중존재유리DNA적이론，채용자주작위고상흡부재체병사용특정설계적시제체계급제취류정，건립일충간편、고효제취뇨액양본중유리갑기화DNA적방법，평개기작위용우뇨액양본적갑기화기인검측기술적가행성。방법사용자주법제취40례성인뇨액중유리갑기화DNA，진행갑기화수식후，자외분광광도계측정DNA적농도화순도。결과결과현시，제취50 mL뇨액가득61~200 ng/μL적갑기화DNA，OD260/280위1.8±0.05。사용갑기화양성대조DNA인물진행PCR급전영후가견목적조대，설명기순도가이만족후속갑기화기인검측화PCR등조작요구。결론뇨액중학실존재유리갑기화DNA，동시자주법제취뇨액유리DNA제취과정간단，차제취순도고，시일충고효적제취방법。
Objective To establish a simple method to extract the methylated ctDNA in urine using magnetic beads as solid phase adsorption carri?er with a specific design reagent system and extraction process,and evaluate its application feasibility for methylated gene detection in urine sample . Methods Fourty cases of methylated ctDNA were extracted in urine using magnetic beads. After methylated modification,the concentration and pu?rity of DNA was determined by ultraviolet spectrophotometer. Results The extraction of 50 mL urine can gain 61?200 ng/mL methylated ctDNA, and the OD260/280 was 1.8 ± 0.05. Conclusion There are methylated ctDNA exist in the urine which can be extracted by magnetic beads. The estab?lished extraction method is simple,effective,and with high purity.