血清抗碳酸酐酶Ⅲ抗体ELISA检测方法的建立与初步应用

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。

Establishment and preliminary application of the ELISA method for anti-carbonic anhydrase III antibody detection

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 erythematosus,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.