肾组织石蜡切片直接免疫荧光染色方法的再改良

Further Improvement in Method of Direct Immunofluorescence Staining of Paraffin-embedded Renal Sections

目的探讨肾组织石蜡切片直接免疫荧光染色再改良的方法,以提高石蜡切片的诊断准确率。方法选择2010年6-12月在北京大学深圳医院行肾组织活检确诊为肾小球疾病的患者44例,其中IgA肾病20例,狼疮性肾炎12例,膜性肾病12例。对其肾组织除常规冰冻切片直接免疫荧光染色外,对石蜡切片进行高温、高压修复和胃蛋白酶消化处理,再分别行免疫球蛋白(IgA、IgG、IgM)、补体(C3、C1q)、纤维蛋白(Fib)、HBsAg及HBcAg直接免疫荧光染色,比较石蜡与冰冻切片的直接免疫荧光染色结果。结果冰冻切片:肾小球数1～12个,平均5个;石蜡切片:肾小球数7～30个,平均15个。20例IgA肾病、12例狼疮性肾炎、12例膜性肾病患者的肾组织行冰冻(IgA、IgG、IgM、C3、C1q、Fib、HbcAg和HbsAg)直接免疫荧光染色时在不同荧光强度中所占比例与行石蜡直接免疫荧光染色比较差异均无统计学意义(均P>0.05)。结论再改良的肾组织石蜡切片直接免疫荧光染色是冰冻切片失败时较好的补救手段。其石蜡切片组织结构更清晰,免疫复合物沉积部位较冰冻切片更易判断。

Objective To improve the method of direct immunofluorescence staining of paraffin-embedded renal sections,and to increase the diagnostic accuracy of paraffin sections.Methods Conventional frozen sections were processed for direct immunofluorescence staining in 44patients（20 cases of IgA nephropathy,12 cases of lupus nephritis and 12 cases of membranous nephropa-thy） who underwent renal biopsy for diagnosis of glomerular disease in Shenzhen hospital of Peking university between June 2010and December 2010.In addition,paraffin-embedded renal sections were exposed to high temperature,high pressure and pepsin,and were examined by direct immunofluorescence staining for immunoglobulins（IgA,IgG and IgM）,complement components（C3,C1q）,fibrin（Fib）,HBsAg and HBcAg.Results of immunofluorescence staining were compared between paraffin sections and frozen sections.Results In frozen sections,the number of glomeruli ranged from 1 to 12（average,5）.In paraffin sections,the number of glomeruli ranged from 7 to 30（average,15）.There were no significant differences in the fluorescence intensity of IgA,IgG,IgM,C3,C1q,Fib,HBcAg and HBsAg between paraffin sections and frozen sections（all P0.05）.Conclusion The further improved method of direct immunofluorescence staining of paraffin-embedded renal sections is an effective means of redress when frozen-section examination fails in patients with glomerular disease.Compared with frozen sections,paraffin sections canclearly show organizational structure and immune-complex deposition site can be easily judged.