血清抗碳酸酐酶Ⅲ抗体ELISA检测方法的建立与初步应用

摘要

目的 建立人血清抗碳酸酐酶(CA)Ⅲ抗体的ELISA检测方法,并对系统性红斑狼疮、皮肌炎、糖尿病肾病、高血压肾病患者和健康人群的血清抗CAⅢ抗体水平进行初步调查.方法 使用抗CAⅢ抗体标准品、CAⅢ及相应酶标抗体建立血清抗CAⅢ抗体ELISA检测方法,验证试剂稳定性、标本保存稳定性,并进行精密度、灵敏度、回收率、抗干扰性等方法学评价；各项技术指标均合格后对系统性红斑狼疮、皮肌炎、糖尿病肾病和高血压肾病患者的血清进行抗CAⅢ抗体水平检测.结果 成功建立ELISA检测人血清抗CAⅢ抗体的方法,其批内精密度为6.2％,批间精密度为8.2％,灵敏度为0.025,回收率为106％,且具有较好的抗干扰性、试剂稳定性和标本保存稳定性.系统性红斑狼疮和糖尿病肾病患者的血清抗CAⅢ抗体水平高于健康对照相(P＜0.05),阳性率分别为43％和18％.皮肌炎和高血压肾病患者的血清抗CAⅢ抗体水平与健康对照组比较无统计学差异(P＞0.05),且未出现阳性结果.结论 使用现有市售试剂进行人血清抗CAⅢ抗体的ELISA检测是可行的,抗CAⅢ抗体可能参与了系统性红斑狼疮和糖尿病肾病的发生发展.%Objective To establish an ELISA method for anti-carbonic anhydrase III (CA III ) antibody detection, and to evaluate the serum level of anti-CA III antibody in normal control group and patients with systemic lupus erythemato-sus, dermatomyositis, 2-type diabetic nephropathy and hypertensive nephropathy respectively. Methods To establish the ELISA method using CA III, anti-CA III antibody and enzyme labeled secondary antibody. To evaluate the stability of the regent and sample, and the sensitivity, stability and anti-interference performance of the ELISA method. To investigate the anti-CA III antibody level in serum of normal control group and patients with systemic lupus erythematosus, dermatomyositis, 2-type diabetic nephropathy and hypertensive nephropathy respectively. Results It was possible to build a method of ELISA for detecting anti-CA III antibody in serum, the sensitivity was 0.025, within-run precision was 6.2% , between-run precision was 8.2% , recovery rate was 106% , and with favorable anti-interference performance, regent stability and sample stability. The level of serum anti-CA III antibody was significantly higher in patients with systemic lupus erythematosus and 2-type diabetic nephropathy compared with the normal control (P <0. 05) , the positive rate was 43% and 18% respectively. The level of serum anti-CA III antibody was similar to the normal control in patients with dermatomyositis and hypertensive nephropathy (P>0. 05) , and there was no postive case. Conclusions It was possible to build an ELISA method for anti-CA III antibody detection using available regents. It was demonstrated that anti-CA III antibody might play a role in the occurrence and development of systemic lupus erythematosus and 2-type diabetic nephropathy.