Dong Hong-rui, Wang Yan-yan, Wang Guo-qin, Sun Li-jun, Cheng Hong, Chen Yi-pu
Department of Nephrology,Beijing Anzhen Hospital,Capital Medical University,Beijing 100029,China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2015 Oct;37(5):562-6. doi: 10.3881/j.issn.1000-503X.2015.05.012.
To evaluate the application of immunohistochemistry and fluorescence staining method in the detection of phospholipase A2 receptor (PLA2R) on paraffin section of renal biopsy tissue,and to find an accurate and fast method for the detection of PLA2R in renal tissue.
The PLA2R of 193 cases were detected by immunohistochemical staining,and the antigen was repaired by the method of high pressure cooker (HPC) hot repair plus trypsin repair. The 193 samples including 139 cases of idiopathic membranous nephropathy (IMN), 15 cases of membranous lupus nephritis, 8 cases of hepatitis B virus associated membranous nephropathy, 18 cases of IgA nephropathy, and 13 cases of minimal change diseases. To compare the dyeing effects, 22 paraffin sections of renal biopsy tissue of IMN cases with positive PLA2R were stained by using 4 different.
of antigen repairing,which included HPC hot repair, HPC hot repair plus trypsin repair, water bath heat repair, and water bath heat repair plus trypsin repair. To compare the dyeing effects, 15 paraffin sections of renal biopsy tissue of IMN cases with positive PLA2R were stained by using 3 different.
of antigen repairing,which included water bath heat repair plus trypsin repair, protease K digestion repair, and pepsin digestion repair.
In 193 cases, the positive rate of PLA2R in IMN cases was 90.6% (126/139), and the other 54 patients without IMN were negative. Twenty-two IMN patients were positive for PLA2R by using the HPC heat repair plus trypsin repaire or the water bath heat repair plus trypsin repair;while only a few cases of 22 IMN cases were positive by using the HPC hot repair alone or water bath heat repair alone. Fifteen IMN patients were positive for PLA2R by using water bath heat repair plus trypsin repair,protease K digestion repair,and pepsin digestion repair, but the distribution of positive deposits and the background were different.
PLA2R immunohistochemical staining can effectively identify IMN and secondary MN. For immunohistochemical staining and immunofluorescence staining, the preferred method of antigen repair is water bath heat repair plus trypsin repair.
评估免疫组织化学和荧光染色法在肾活检组织石蜡切片中检测磷脂酶A2受体（PLA2R）的应用，寻找一种准确、快速检测肾组织中PLA2R的方法。
采用免疫组织化学染色法检测193例患者的PLA2R，抗原修复采用高压锅热修复加胰蛋白酶修复法。193份样本包括139例特发性膜性肾病（IMN）、15例膜性狼疮性肾炎、8例乙型肝炎病毒相关性膜性肾病、18例IgA肾病和13例微小病变病。为比较染色效果，对22例PLA2R阳性的IMN患者肾活检组织石蜡切片采用4种不同的抗原修复方法进行染色，包括高压锅热修复、高压锅热修复加胰蛋白酶修复、水浴热修复、水浴热修复加胰蛋白酶修复。为比较染色效果，对15例PLA2R阳性的IMN患者肾活检组织石蜡切片采用3种不同的抗原修复方法进行染色，包括水浴热修复加胰蛋白酶修复、蛋白酶K消化修复、胃蛋白酶消化修复。
193例中，IMN患者PLA2R阳性率为90.6%（126/139），其余54例非IMN患者为阴性。22例IMN患者采用高压锅热修复加胰蛋白酶修复或水浴热修复加胰蛋白酶修复时PLA2R呈阳性；而单独采用高压锅热修复或水浴热修复时，22例IMN患者中只有少数病例呈阳性。15例IMN患者采用水浴热修复加胰蛋白酶修复、蛋白酶K消化修复、胃蛋白酶消化修复时PLA2R呈阳性，但阳性沉积物分布及背景不同。
PLA2R免疫组织化学染色可有效鉴别IMN和继发性MN。对于免疫组织化学染色和免疫荧光染色，首选的抗原修复方法是水浴热修复加胰蛋白酶修复。
