肾活检冰冻切片不同储存条件对免疫荧光结果的影响

Effect of different storage temperatures on immunofluorescence results in frozen sections of kidney biopsies

目的：探讨肾活检组织冰冻切片储存温度对免疫荧光结果的影响。方法：选择2015年1月～2015年6月浙江省人民医院肾脏病科肾脏活检标本100例,将同一肾穿组织同时切两套切片分别储存于-70℃低温冰箱和-12℃冷冻切片机内,其中储存于-70℃低温冰箱的切片作为实验组,储存于-12℃冷冻切片机内的切片作为对照组,次日用直接免疫荧光法检测免疫球蛋白（IgG、IgA和IgM）、补体（C3、C4和C1q）及纤维蛋白原,间接免疫荧光法检测Ⅳ型胶原a3和a5链,荧光显微镜下观察定性及半定量结果。结果：实验组3＋～4＋IgG 16例,对应的对照组IgG均为1＋～2＋,阳性强度明显弱于实验组,差异有统计学显著性（P〈0.05）;实验组3＋～4＋a3 99例,对照组1＋～2＋a3 92例,阴性8例;实验组3＋～4＋a5 99例,对照组1＋～2＋a5 92例,阴性8例,对照组阳性强度减弱,差异有统计学显著性（P〈0.05）。实验组4＋IgA 15例,3＋IgA 19例,3＋IgM 15例,2＋IgM11例,4＋C3 13例,3＋C3 12例,相对应的对照组阳性强度没有明显减弱,差异无统计学显著性。结论：肾活检冰冻切片免疫荧光结果受切片储存温度影响大,其中对IgG和Ⅳ型胶原免疫荧光阳性强度影响更大,建议肾脏活检组织冰冻切片置于-70℃低温冰箱储存。

AIM： To investigate the effect of storage temperature in renal biopsy tissue frozen section on immunofluorescence results. METHODS： One hundred renal biopsy samples of Zhejiang Province People＇s Hospital from January to June,2015 were enrolled. Two sets of cutting slices were stored in- 70 ℃ low temperature refrigerator as experimental group and in- 12 ℃ freezing cryostat as control group. The immunoglobulins（ IgG,IgA and IgM） and complements（ C3,C4 and C1q） as well as fibrinogen were detected with direct immunofluorescence in the next day. The type Ⅳcollagen a3 and a5 chains were also detected by indirect immunofluorescence,and the qualitative and semi-quantitative results were observed under the fluorescence microscope. RESULTS： There were 16 cases of 3 ＋ ～ 4 ＋ IgG in the experimental group,while the corresponding IgG was all 1 ＋ ～ 2 ＋ in control group,significantly weaker than that in experimental group. There were 99 cases of 3 ＋ ～ 4 ＋ a3 and 3 ＋ ～ 4 ＋ a5 in experimental group,white there were 92 cases of 1 ＋～ 2 ＋ a3/a5 and 8 cases negative in control group. The positive intensity was decreased in control group with statistical difference between the 2 groups（ P〈0. 05）. There were 15 cases of 4 ＋ IgA,19 cases of 3 ＋ IgA,15 cases of 3 ＋ IgM,11 cases of 2 ＋ IgM,13 cases of 4 ＋ C3,and 12 cases of 3 ＋ C3 in experimental group. The control group were similar to the results of experimental group,and no difference between the 2 groups was observed. CONCLUSION： The immunofluorescence results of renal biopsy frozen sections are highly affected by the section storage temperature,which has greater influence on the immunofluorescence positive intensity of IgG and type Ⅳ collagen. The renal biopsy frozen section should be stored in- 70 ℃ low temperature refrigerator.