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机构地区:[1]南方医科大学南方医院肾脏病实验室,广东广州510515
出 处:《南方医科大学学报》2013年第10期1448-1452,共5页Journal of Southern Medical University
基 金:国家自然科学基金青年基金(81200502)~~
摘 要:目的分析3种特殊染色及免疫染色方法对区分肾淀粉样变类型的准确性、灵敏度和着色强度的影响,以提高临床的诊治水平。方法选择30例肾淀粉样变患者的病理切片,采用3种酸化刚果红染色和5种抗原修复法,对轻链蛋白(κ、λ)、淀粉样蛋白A等指标进行冰冻组织及石蜡组织的免疫荧光、免疫组化染色。使用IPP图像分析系统进行半定量分析及荧光着色半定量分析,观察比较阳性部位着色的准确性、灵敏度和着色强度效果。结果酸化甲醇刚果红染色法优于酸化Bennhold's刚果红染色法、酸化Alkaland's刚果红染色法,IPP半定量分析冰冻组织的免疫荧光、免疫组化优于石蜡组织(P=0.02)。荧光半定量分析着色强度相差一个+左右;石蜡组织暴露抗原的方法中蛋白酶K消化优于抗原热修复、胰酶或胃酶消化(P=0.01)。结论酸化甲醇刚果红染色法在肾组织淀粉样变部位的阳性着色准确性、灵敏度、着色强度和重复率均优于Bennhold's刚果红染色法、酸化Alkaland's刚果红染色法。起到初步分型及辅助验证免疫荧光染色结果的作用;进行冰冻组织轻链蛋白(κ、λ)及淀粉样蛋白A免疫荧光染色,更有利于肾淀粉样变的准确分型,如果无冰冻组织也可选用蛋白酶K消化石蜡组织免疫荧光染色。Objective To analyze the effect of special staining and immunohistochemical staining in distinguishing the types of renal amyloidosis to improve the diagnosis accuracy. Method Congo red staining with different methods, and immunohistochemical staining of Kappa, Lambda and Amyloid A with different antigen retrieval methods were used for staining frozen and paraffin-embedded renal tissue sections. Results Wright's Congo red staining produced a better contrast and a higher resolution and showed a greater stability than the other 2 methods after repeated use for staining the renal tissue sections (P〈0.05). Immunofluorescent staining produced better results in frozen renal tissue sections than in paraffin-embedded tissues. Immunofluorescent staining produced had better performance than immunohistochemical staining in paraffin-embedded tissues. The retrieval effect with protein kinase K was the best among the antigen retrieval methods in paraffin-embedded tissues. Conclusion Wright's Congo red staining is better than the other 2 methods in diagnosing renal amyloidosis. Immunohistochemical staining of Kappa, Lambda and Amyloid A in frozen renal tissue sections is necessary to discriminate the types of renal amyloidosis. For paraffin-embedded renal tissues, antigen retrieval using protein kinase K is better than the other 2 methods.
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