HIV感染者的自身抗体检测分析

目的分析人免疫缺陷病毒（HIV）感染者中出现的自身抗体和自身免疫现象。方法回顾性分析经间接免疫荧光法检测过自身抗体的HIV感染者的临床和相关实验室指标。结果共有19例HIV感染者检测了自身抗体,明确合并银屑病、溃疡性结肠炎、特发性血小板减少性紫癜（ITP）的患者各1例;8例出现风湿性症状,主要表现为原因不明的皮疹（5例）、关节疼痛（2例）和肌痛（1例）。10例自身抗体检测结果阳性,出现的自身抗体分别为抗核抗体（ANA）、抗平滑肌抗体（SMA）、抗线粒体抗体（AMA）、抗细胞骨架抗体（CS）、抗胃壁细胞抗体（AP—CA）。自身抗体阳性组患者的IgG和IgA水平明显高于自身抗体阴性组（P〈0．05）。结论HIV感染者中可检测出自身抗体,而且可以合并有自身免疫病或免疫亢进的现象。     

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

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.     

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

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.     

药物性肝损伤患者的自身抗体分析

目的分析药物性肝损伤患者出现的自身抗体。方法对3000例经间接免疫荧光法及免疫印迹法进行过自身抗体检测的肝功能异常患者进行回顾分析。结果3000例肝功能异常患者中药物性肝损伤患者36例,占肝功能异常患者的1.2‰。36例药物性肝损伤患者中有29例自身抗体检测阳性,其阳性率分别为抗核抗体(ANA)66.7%(24/36)、抗平滑肌抗体(SMA)16.7%(6/36)、抗线粒体抗体(AMA)19.4%(7/36)、抗肝脏特异蛋白抗体(LSP)2.8%(1/36)、抗细胞骨架抗体(CS)19.4%(7/36)、抗胃壁细胞抗体(APCA)5.6%(2/36)、抗心肌抗体(AHRA)8.3%(3/36)。ANA、SMA和AMA阳性的拟诊药物性肝损伤患者进一步检测AMA亚型-M2、抗肝肾微粒体1(LKM-1)和抗可溶性肝抗原/肝胰抗原(SLA/LP),结果全部阴性;肝细胞型和胆汁淤积型、混合型自身抗体检测结果比较,其阳性率分别为95.2%(20/21),60.0%(9/15)。自身抗体阳性的药物性肝损伤患者其免疫球蛋白升高的比例比自身抗体阴性的要高,两者之间的差异有统计学意义(P<0.05)。结论药物性肝损伤患者血清中存在多种自身抗体但滴度较低;自身抗体(尤其是ANA、SMA和AMA)阳性的肝功能异常患者在拟诊药物性肝损伤时要注意自身免疫性肝脏疾病的排除。     

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

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.     

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

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.     

3-磷酸甘油酸脱氢酶及其抗体对自身免疫性肝炎的诊断意义

目的 克隆D-3-磷酸甘油酸脱氢酶(Phgdh)的cDNA,构建重组表达质粒,纯化重组蛋白,初步探讨Phgdh抗体在自身免疫性肝炎诊断中的意义. 方法血清蛋白质组学方法鉴定差异蛋白,构建重组表达载体,在大肠杆菌中表达,融合蛋白经Ni-NTA树脂柱亲和层析纯化,通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳及Western blot法进行免疫鉴定;应用表达蛋白建立间接酶联免疫吸附法,检测60名健康体检者、65例自身免疫性肝炎(AIH)、122例原发性胆汁性肝硬化、56例慢性乙型肝炎,117例慢性丙型肝炎患者血清中抗Phgdh抗体.组间率的比较用χ2检验.结果 经重组质粒测序和酶切鉴定结果证实,Phgdh目的 基因已正确插入原核表达载体中,基因序列正确,符合表达框架.十二烷基硫酸钠聚丙烯酰胺凝胶电泳检测表达产物在相对分子质量约6.0×104附近有一明显的蛋白表达带,Western blot分析结果显示重组蛋白具有Phgdh抗原反应性.酶联免疫吸附法检测血清标本结果显示,AIH、原发性胆汁性肝硬化、慢性乙型肝炎、慢性丙型肝炎及正常人抗Phgdh抗体阳性检出率分别为66.15%、21.42%、12.50%、6.83%和3.30%.AIH组Phgdh自身抗体阳性率与疾病对照组及正常对照组比较,差异均具有统计学意义(P<0.01).结论 本研究成功克隆了Phgdh的cDNA,并将其在大肠杆菌中成功表达.该抗体主要在AIH患者中检出,此抗体的检测有助于提高AIH的临床诊断水平.     

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

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.     

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.     

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

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.