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中文摘要：
目的建立从肾穿刺活检石蜡组织切片提取DNA并进行PCR扩增的方法。方法肾脏病理诊断为乙肝病毒相关性肾小球肾炎、原发性IgA肾病和膜性肾病各2例的肾活检组织蜡块，包埋时间1个月-5年。提取方法包括清洁切片、融蜡、消化及DNA的提取。紫外分光光度计测定DNA浓度。应用聚合酶链反应（PCR）扩增血管紧张素转换酶（ACE）和乙型肝炎病毒核心抗原（HBcAg）基因片段。结果2张2Ixm厚石蜡切片获取的DNA总量为10-30μg。ACE基因的PCR扩增结果显示，6例标本均在490bp和（或）190bp处显示清晰条带。HBcAg的PCR扩增结果显示，2例乙肝病毒相关性肾小球肾炎标本在132bp处显示清晰条带，其余4例阴性。结论本方法可以从肾穿刺活检石蜡切片中获取足够高质量DNA并进行PCR扩增，方法简便，结果稳定，不受包埋时间长短的限制，为肾脏疾病的辅助诊断、肾活检组织的大宗遗传学分析提供了可能。
英文摘要：
Objective To establish a method for DNA extraction and PCR amplification from renal biopsy paraffin sections. Methods Six paraffin embedded renal biopsy tissue blocks, which pathological diagnoses were hepatitis B associated glomerulonephritis （2 case）, primary IgA nephropathy（2 case） and membrane nephropathy（2 case）, were employed. Two or three sections at 2 μm thickness were cleanly cut and placed on microscope slides, Sections were incubated at 70℃ for 30 min to melt down paraffin and deparaffinized with three changes of xylene, Digestion buffer was freshly made by mixing 150 μl of protein K solution （10 mg/ml） with 850 μl of a mixture of the followings： 0.5 ml Tween 20, 0,2 ml 0,5 mol/L EDTA（pH 8.0）, 5,0 ml 1 mol/L Tris（pH 8.5）, 94,3 ml of distilled water, Tissues on the slides were removed by 10 μ1 Eppendoff pipette tip and each sample was placed in a separate tube. Samples were digested for 48 h and DNA concentration was measured, Angiotensin converting enzyme gene and HBcAg gene were amplified by PCR. Results DNA extraction of samples ranged from 10 μg to 30 μg. ACE gene PCR amplification results showed all samples displayed 490 bp and/or 190 bp lanes, HBcAg gene PCR amplification results showed the two hepatitis B associated glomerulonephritis samples displayed 132 bp lane while the primary IgA nephropathy and membrane nephropathy samples were negative,Conclusion This method is able to obtain comparable quality and quantity of DNA extracts which yields uniform PCR products from renal biopsy paraffin sections and provides the possibility for large genetics analysis of renal biopsy tissues.
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