抗乙醇脱氢酶抗体ELISA法的建立及其对自身免疫性肝炎诊断的价值

目的 建立血清抗乙醇脱氢酶(alcohol dehydrogenase,ADH)抗体ELISA法,并评价抗.ADH在诊断自身免疫性肝炎(AIH)中的价值.方法 用免疫印迹试验对酵母ADH与人血清抗.ADH之间的反应性进行验证.用酵母ADH建立检测血清抗-ADH的ELISA法.以67例AIH、94例原发性胆汁性肝硬化(PBC)、199例慢性乙型肝炎(CHB)、132例慢性丙型肝炎(CHC)、24例酒精性肝病(ALD)和99例结缔组织病(CTD)患者及31名健康对照者为研究对象,对其血清中的抗-ADH进行检测并统计其阳性率,阳性率的比较用χ2检验.结果 建立了一种检测人血清抗-ADH的ELISA法,并确定了最佳反应条件;免疫印迹试验证实酵母ADH与人血清抗-ADH有良好反应性.AIH患者血清抗-ADH阳性率为59.7%(40/67),高于健康对照组(0,χ2=31.271,P<0.05)、PBC组(6.4%,χ2=54.492,P<0.05)、CHB组(14.1%,χ2=54.848,P<0.05)、CHC组(21.2%,χ2=29.269,P<0.05)、ALD组(25.0%,χ2=8.512,P<0.05)和CTD组(43.4%,χ2=4.229.P<0.05).结论 AIH患者血清中抗-ADH阳性率高于其他肝病和某些自身免疫性疾病患者,可能对AIH有一定辅助诊断价值.

Establishment of an enzyme-linked immunosorbent assay for the detection of alcohol dehydrogenase autoantibody and the evaluation of its usage in the diagnosis of autoimmune hepatitis

Objective To establish the enzyme-linked immunosorbent assay (ELISA) for the detection of serum alcohol dehydrogenase (ADH) antibody and evaluate its role in its diagnosis of autoimmune hepatitis( AIH ). Methods The reactivity between yeast ADH and human anti-ADH serum antibody was tested by Western blot analysis. ELISA was established using yeast ADH. The method was applied in serums of 67 AIH patients,94 primary biliary cirrhosis(PBC) patients, 199 chronic hepatitis B (CHB) patients, 132 chronic hepatitis(CHC) patients, 24 alcohol hepatitis disease(ALD) patients, 99 connective tissue disease(CTD) patients and 31 healthy individuals. The positive rate of ADH antibody in the patients and healthy individuals was measured. The χ2 test was used to compare the positive rates. Results The ELISA method for detecting human anti-ADH serum antibody was established successfully and the optimum reaction conditions were defined. Western blot showed that yeast ADH has cross reactivity with human anti-ADH antibody. The positive rate of anti-ADH antibody in the AIH group 59. 7% ,40/67 ] was higher than that in the normal control group(0,χ2 = 31. 271 ,P <0. 05), PBC group (6. 4% ,χ2 =54. 492,P <0. 05), CHB group( 14. 1% ,χ2 =54. 848,P <0. 05) ,CHC group(21.2% ,χ2 = 29.269,P<0.05), ALl) group ( 25. 0% ,χ2 =8.512,P <0.05)and CTD group ( 43. 4% ,χ2 =4.229, P <0. 05). Conclusions Compared with the PBC, CHB, CHC, ALD and CTD group, the anti-ADH antibody positive rate in the serums of AIH was significantly increased. The antibody may be helpful to the diagnosis of autoimmune hepatitis.