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背景:近年的研究发现在癫痫患者血清中存在各种自身抗体,如抗核抗体、抗心磷脂抗体、抗β2糖蛋白1抗体、抗谷氨酸脱羧酶抗体等,表明癫痫患者存在自身免疫机制。目的:检测癫痫患儿血清抗谷氨酸受体1抗体水平。设计:对照实验。单位:大连大学医学院和吉林化学集团总医院及日本三重大学。对象:选择2002-12/2004-01青岛市妇幼保健医院门诊及住院癫痫患儿56例为患儿组,男30例,女26例;年龄3个月～13岁。选择同期吉林化学集团总医院儿科门诊健康体检患者48例和该院脑外科就诊的脑肿瘤患者12例为对照组,其中男38例,女22例;年龄6个月～17岁。监护人均对检测项目知情同意。方法:谷氨酸受体1多肽抗原包被微量反应板,采用抗原和抗体同时稀释的方法(棋盘滴定法)测定酶联免疫吸附反应检测抗谷氨酸受体1抗体的最适实验条件。用酶联免疫吸附方法测定癫痫患儿血清中谷氨酸受体1抗体。主要观察指标:酶联免疫吸附测定方法检测血清抗谷氨酸受体1抗体水平。结果:谷氨酸受体1多肽抗原包被浓度为20mg/L;辣根过氧化物酶-羊抗人IgG和血清稀释度分别为1∶1500和1∶50;批内和批间变异系数分别为4.48%～8.80%和11.18%～16.60%;特异性吸收试验结果显示吸光度值平均下降2.4倍。患儿组血清中抗谷氨酸受体1抗体的阳性率明显高于对照组(40%,5%,P<0.01)。结论:①酶联免疫吸附法检测血清抗谷氨酸受体1抗体特异性强,稳定性好。②癫痫患儿可能存在抗谷氨酸受体1抗体的自身免疫反应。
BACKGROUND: Recent studies have found that autoantibodies such as antinuclear antibodies, anticardiolipin antibodies, anti-β2 glycoprotein 1 antibodies and anti-glutamic acid decarboxylase antibodies exist in the sera of patients with epilepsy, indicating that there is an autoimmune mechanism in patients with epilepsy . Objective: To detect serum anti-glutamate receptor 1 antibody level in children with epilepsy. Design: Control experiment. Unit: Dalian University Medical College and Jilin Chemical Group General Hospital and Japan’s Mie University. PARTICIPANTS: 56 patients with outpatients and inpatients with epilepsy from March 2002 to January 2004 in Qingdao MCH hospital were selected as the pediatric group, 30 males and 26 females, aged 3 months to 13 years old. Select the same period, Jilin Chemical Group General Hospital pediatric outpatient checkup 48 patients and brain surgery in the hospital 12 patients with brain tumor control group, 38 males and 22 females; aged 6 months to 17 years. Guardian informed consent of the test items. Methods: The glutamate receptor 1 polypeptide antigen was coated on the microtiter plate, and the optimum conditions for detecting anti-glutamate receptor 1 antibody by enzyme-linked immunosorbent assay (ELISA) were determined by the method of simultaneous dilution of antigen and antibody (checkerboard titration). Serum glutamate receptor 1 antibody in children with epilepsy was detected by enzyme-linked immunosorbent assay. MAIN OUTCOME MEASURES: Serum anti-glutamate receptor 1 antibody levels were measured by enzyme linked immunosorbent assay. Results: The concentration of glutamate receptor 1 polypeptide antigen was 20 mg / L; the HRP-goat anti-human IgG and serum dilution were 1:1500 and 1:50 respectively; Respectively, 4.48% ~ 8.80% and 11.18% ~ 16.60%. The results of specific absorption test showed that the absorbance decreased by an average of 2.4 times. The positive rate of anti-glutamate receptor 1 antibody in children’s serum was significantly higher than that of the control group (40%, 5%, P <0.01). Conclusion: ① The serum anti-glutamate receptor 1 antibody was detected by enzyme-linked immunosorbent assay (ELISA) with high specificity and good stability. ② children with epilepsy may exist anti-glutamate receptor 1 antibody autoimmune response.