ANA和ENA检测在自身免疫性疾病中的应用评价

探讨抗核抗体和抗可溶性核抗原抗体的不同检测方法对自身免疫性疾病诊断的指导意义。回顾性分析2007年1月至2009年10月间在长征医院就诊的患者血清自身抗体的检测结果,549位患者,其中自身免疫病患者224例,非自身免疫病325例,所有患者血清同时检测ANA和ENA。以Hep-2细胞/肝组织为基质的间接免疫荧光法检测ANA,免疫印迹法检测ENA。两种检测方法产生4种检出模式:ANA+/ENA+、ANA-/ENA-、ANA+/ENA-和ANA-/ENA+。前两种模式共占检测的62.84%。ANA和ENA在自身免疫病患者中的阳性率(63.4%和58.5%)显著高于非自免病患者(16.9%和25.2%),ANA和ENA在自身免疫病组和非自身免疫病组间阳性率比较,差异有统计学意义(P<0.01)。两检测结果仅在MCTD和SLE患者中存在相关性(P<0.01),在其他观察组中不存在相关性(P>0.05)。间接免疫荧光法相对费时,而且需要操作者具备一定的经验,作为初筛实验,ANA的检测较ENA有更高的灵敏度,两者联合检测则将有利于提高检测的灵敏度和可靠性。     

抗核抗体与抗核抗体谱联合检测在自身免疫性疾病诊断上的应用

目的探讨抗核抗体(antinuclear antibody,ANA)和抗核抗体谱(antinuclear antibodies spectrum,ANAs)联合检测在自身免疫性疾病(autoimmune diseases,AID)诊断上的应用方案及价值,从而降低现症及潜在AID患者的漏检率。方法对1 231份本院门诊和住院疑似AID患者的血清标本分别采用间接免疫荧光法(indirect immunofluorescence,IIF)和免疫印迹法(immumoblottest,IBT)检测ANA和ANAs,对其中IIF-ANA1∶100(-)/IBT-ANAs(+)标本分别进行1∶10及1∶32稀释后采用IIF检测ANA,最后对上述检测结果进行统计分析。结果 1 231份血清标本中ANA阳性414份,占33.63%,ANAs阳性429份,占34.85%。按不同检测方案进行分组,发现同时检测IIF-ANA与IBT-ANAs的方案对确诊/疑似AID病人检出率最高(43.05%),分别与应用IIF-ANA进行AID的初筛的方案、仅检测IBT-ANAs的方案相比均有统计学意义(χ2=23.12,P<0.05,χ2=17.42,P<0.05)。对116份(9.42%)IIF-ANA 1∶100(-)/IBT-ANAs(+)的标本分别进行1∶32及1∶10稀释后再采用IIF检测ANA,发现其中103份标本ANA荧光结果≥1∶32,10份标本ANA荧光结果<1∶32而≥1∶10,3份标本ANA荧光结果<1∶10;将标本增加1∶32和1∶10稀释度可将AID确诊/疑似病例检出率分别提高8.37%(χ2=20.84,P<0.05)与9.18%(χ2=24.70,P<0.05),但二者之间相比则无统计学意义(χ2=0.17,P>0.5)。结论 IIF-ANA可应用于大范围AID病人初筛以减轻病人经济负担及实验室工作量压力。但是仅检测IIF-ANA或IBT-ANAs均可导致临床上患有AID或潜在的AID病人的漏检。联合检测IIF-ANA和IBT-ANAs,尤其是对临床高度怀疑AID、且ANA荧光结果≥1:32的标本进行IBT-ANAs的检测可显著提高检出率,从而降低现症及潜在AID患者的漏检率。     

抗角蛋白抗体及相关自身抗体在RA诊断中的应用评价

目的 评价抗角蛋白抗体（AKA）及其相关自身抗体对诊断类风湿关节炎（RA）的敏感性和特异性以及在临床上的意义。方法 收集血清279例，包括正常人30例，RA56例，SLE114例，其它结缔组织病179例，检测AKA（IFA）、RF、ANA及ssDNA。结果 4种自身抗体对诊断RA疾病的敏感性和特异性为：AKA39．3％和96．0％；RF37．5％和85．5％；ANA55．4％和39．5％；ssDNA37．5％和49．8％；敏感性无显著性差异（P＞0．05），特异性则AKA明显高于其它3种抗体（P＜0．001）。RF阳性率在RA患者AKA阳性组和阴性组有显著性差异（P＜0．001），且RF含量与AKA滴度呈正相关；而ssDNA和ANN则无显著性差异（P＞0．05）。结论 AKA是RA的又一个特异性抗体，联合检测AKA及RF对于RA的诊断与早期诊断具有重要意义。     

免疫荧光法在结直肠肿瘤自身抗核抗体检测中的临床价值

目的 探讨免疫荧光法在结直肠肿瘤自身抗核抗体检测中的临床价值.方法 选取我院2015年1月至2016年1月85例结直肠癌患者为恶性肿瘤组,85例结直肠良性肿瘤患者为良性肿瘤组,85例健康体检者为对照组.采用间接免疫荧光法对受试者的抗核抗体进行检测,对比3组抗核抗体检测及在细胞内荧光定位的结果.结果 ①恶性肿瘤组中转移组抗核抗体阳性率和无转移组阳性率比较无明显差异(P＞0.05);恶性肿瘤组抗核抗体阳性率明显高于良性肿瘤组和对照组x2=16.940、37.531,P=0.000、0.000;良性肿瘤组抗核抗体阳性率高于对照组,x2=5.328,P=0.021.②恶性肿瘤组和良性肿瘤组、对照组抗胞质抗体荧光阳性率无明显差异(P＞0.05).和对照组比较,恶性肿瘤组、良性肿瘤组抗核抗体荧光阳性率明显升高x2=5.221、20.299,P=0.022、0.000;恶性肿瘤抗胞质抗体荧光阳性率明显升高,x2=13.523,P =0.000.和良性肿瘤组比较,恶性肿瘤组抗核抗体和抗胞质抗体荧光阳性率明显升高,x2=5.842、8.947,P=0.016、0.003.③恶性肿瘤转移组和无转移组抗核抗体和抗胞质抗体荧光阳性率无明显的差异,不具有统计学意义(P＞0.05).结论 采用免疫荧光法对消化系统恶性肿瘤患者的自身抗核抗体进行检测具有重要的意义,能够有效地判断肿瘤的病情和治疗效果,为临床免疫性治疗提供一定的依据.     

抗核抗体谱检测在诊断系统性红斑狼疮中的意义

探讨抗核抗体(ANA)和抗核抗体谱(ANAs)检测对诊断系统性红斑狼疮(SLE)患者的临床意义。用间接免疫荧光法(IIF)和免疫印迹法检测106例SLE组和30名对照组血清ANA及ANAs中的12种抗体。结果表明:SLE组ANA阳性率为86.8%,ANAs中抗ds-DNA、抗Scl-70、抗Jo-1、抗nRNP、抗Sm、抗SS-A、抗SS-B、抗Ro-52、抗CENP-B、抗AnuA、抗AHA和抗核糖体P蛋白的阳性率分别为28.3%、0.9%、0.9%、35.9%、17.0%、39.6%、18.9%、41.5%、9.4%、29.3%、31.1%和9.4%。ANA和ANAs联合检测提高了诊断的敏感性,对SLE的诊断和治疗有重要意义。     

间接ELISA检测抗核抗体及其临床应用

用小鼠肝细胞核包被聚苯乙烯反应板建立了间接ＥＬＩＳＡ法检测抗核抗体。通过特异性、精密度试验，达到方法学要求。对临床５１例病人检测，与间接荧光抗体法比较，两种方法检出率无明显差异（ｘ２＝３．６７Ｐ＞０．０５）。可进行半定量分析及大批量操作。     

普鲁卡因酰胺等诱导BALB／c小鼠产生抗核抗体

为了建立药物诱导狼疮（DIL）的动物模型,用普鲁卡因酰胺（Pca)、盐酸肼苯哒嗪（Hyd)和2,2-偶氮-双（3-乙苯基-噻唑啉-6-硫酸）（ABTS）对不同品系小鼠腹腔给药,用ELISA和间接免疫荧光法检测抗核抗体。三种药物都能够诱导小鼠产生抗核抗体,Pca诱导小鼠产生核抗体的阳性率为25%,而Hyd、ABTS诱导的阳性率均为12.5%。Pca、Hyd和ABTS都能诱导BALB/c小鼠而不是C57BL/6小鼠产生抗核抗体,其中DNA特异的抗体主要是IgG,而对组蛋白特异的抗体主要是IgM。结果表明在人体导致DIL的药物Pca、Hyd和ABTS能诱导昆明鼠及BALB/c小鼠产生抗核抗体,提示DIL的发生受遗传背景的影响。     

抗核抗体谱检测的临床诊断意义

目的:分析抗核抗体线性免疫印迹分析法( ANA-LIA)和抗核抗体间接免疫荧光法在自身免疫性疾病临床的诊断价值.方法:ANA-LIA使用德国IMTEC公司IMTEC-Human GmbH抗核抗体谱试剂检测;抗核抗体用间接免疫荧光法检测.结果:用间接免疫荧光法检测抗核抗体300例,用免疫印迹法检测抗核抗体谱263例,ANA荧光法与ANA-LIA免疫印迹法检测阳性结果无显著性差异(P＞0.05);30例同时进行ANA、ANA-LIA检测,结果一致率73.3％;ANA-LIA漏检率20％,错检率6.7％.结论:以间接免疫荧光法检测抗核抗体缺乏特异性,而抗核抗体谱免疫印迹法具有高特异性,但存在着较高的漏检率,因此在临床实际应用中将两种方法联合应用,检测效果更好.     

7865例抗核抗体检测者中致密斑点型核型结果的回顾性分析

目的 探讨致密斑点型抗核抗体(dense fine speckled,DFS)在抗核抗体(antinuclear antibody,ANA)中的检出率及其在类风湿关节炎(rheumatoid arthritis,RA)患者中的临床价值.方法 回顾性分析2017年至2020年本院就诊患者7 865例ANA检测标本,采用间...     

87例抗核抗体斑点型阳性患者血清特异性抗核抗体谱分析

目的探讨血清斑点型抗核抗体（ANA）与特异性抗核抗体谱（ANAS）中不同项目阳性组合之间的相关性。方法分析87例斑点型ANA阳性病例的抗ANAS（抗Ul-nRNP、Sm、SS-A、SS—B、Scl-70、Jo-1、SCL-70、PM—Scl、centromereB、nukleosome、PCNA、ds—DNA、histones、ribosomal—Pprotein、AMA-M2抗体）多肽谱类型。结果斑点型ANA的特异性ANAS不同项目阳性组合类型达到23种,除SCL-70、PM—Scl外,其他13种特异性ANAS成分均见阳性。ds—DNA阴性的histone、Sm、nukleosome、PCNA、Rib-P．P单独阳性或合并其他抗体阳性率高达19．5％（17／87）,联合检测这些抗体能提高系统性红斑狼疮（SLE）诊断的敏感性。ANAS阳性率与ANA滴度之间没有绝对关系。结论不能简单地依据荧光核型来推断抗ANAS抗体的具体类型;多项指标联合检测能提高诊断疾病的敏感性和特异性;斑点型ANA低滴度时也应作特异性ANAS检测。     

Evaluation of Recombinant Antigen-Based Assays for Diagnosis of Bullous Autoimmune Diseases

The diagnosis of autoimmune bullous diseases is based on clinical observation and on the presence of autoantibodies directed to molecules involved in the adhesion systems of the skin. Immunofluorescence assays are the currently accepted method for detection of autoantibodies; such assays depend greatly on the skill of operators and are difficult to standardize. Recombinant desmoglein-1 (Dsg1), Dsg3, and BP180 peptides, the main autoantigens in pemphigus or bullous pemphigoid, have been used to develop new quantitative enzyme immunoassays (EIA) for the detection of specific antibodies. The present study was undertaken to evaluate the sensitivity and specificity of these immunoassays and to determine the correlation between the results and the clinical aspects of diseases. Serum samples from patients with pemphigus vulgaris, pemphigus foliaceus, bullous pemphigoid, or mucous membrane pemphigoid, from healthy individuals, and from patients with unrelated autoimmune conditions were tested. Anti-desmoglein reactivity was detected in all the patients with pemphigus and in none of the controls. Patients with the more benign form of cutaneous disease had anti-Dsg1 antibodies, while patients with deeper cutaneous lesions or with mucosal involvement had anti-Dsg3 reactivity also, or exclusively. The BP180-based assay was positive for 66.6% of patients with bullous pemphigoid and for none of the patients with mucous membrane pemphigoid, and no reactivity was detected in the control sera. In conclusion, the anti-Dsg1 and anti-Dsg3 assays are useful in the diagnosis of pemphigus and provide information on the clinical phenotype of the disease. However, the sensitivity of EIA for detection of autoantibodies in bullous pemphigoid should be improved by the use of additional antigens or epitopes.     

Antibody assays for varicella-zoster virus: comparison of competitive enzyme-linked immunosorbent assay (ELISA), competitive radioimmunoassay (RIA), complement fixation, and indirect immunofluorescence assays

The competitive ELISA test was compared with competitive RIA, complement fixation, and indirect fluorescent antibody test for the detection of antibody to varicella-zoster virus (VZV). ELISA and RIA were the most sensitive tests, being tenfold as sensitive as indirect immunofluorescence and 20 times as sensitive as complement fixation, which was surprisingly inadequate for detecting anti-VZV in sera from regular haemodialysis patients. The ELISA test was found to be the most suitable assay, employing a routine complement fixation test (CFT) grade viral antigen and not requiring radiolabelling and counting facilities, which are unavailable in most virology laboratories.     

原发性胆汁肝硬化患者血清中抗线粒体抗体及其亚型和抗核抗体的联合检测的临床价值

为了分析原发性胆汁性肝硬化(primary biliary cirrhosis,PBC)患者的抗线粒体抗体(anti-mitochondria antibody,AMA)及其M2、M4、M9亚型、抗核抗体(antinuclear antibody,ANA)的阳性率。应用间接免疫荧光法检测ANA、AMA,免疫斑点法检测AMA M2、M4、M9。结果显示91例PBC患者中AMA有89例为阳性。其中96.7%(88/91)的患者M2型阳性,45.1%(41/91)M4型阳性,2.2%(2/91)M9型阳性。39/91例患者ANA阳性,其中21例为核膜型。ANA、AMA及其M2、M4、M9亚型联合检测对于PBC患者的诊断有重要价值。     

ELISA检测梅毒IgM抗体在早期梅毒诊断中的应用

为探讨ELISA法检测梅毒螺旋体IgM抗体（Tp-IgM)在早期梅毒诊断中的意义。本文对确诊的67例早期梅毒、3例早期神经梅毒,3例不定期潜伏梅毒患者与26名非梅毒对照。采用ELISA方法,进行血清/脑脊液的梅毒螺旋体IgM抗体检测,同时作RPR及TPPA检测,结果表明,ELISA,RPR及TPPA的特异性分别为100%、88.5%和100%;ELISA法对于早期梅毒的敏感性（95.5%)高于RPR和TPPA（均为83.6%),而在3例不定期潜伏梅毒血清和3例早期神经梅毒脑脊液中,ELISA各检出2例阳性,提示ELISA法检测Tp-IgM对早期梅毒的诊断价值优于RPR和TPPA,对早期神经梅毒和不定期潜伏梅毒的诊断不具优势。     

TPO-Ab在甲状腺自身免疫性疾病中的应用

目的：探讨甲状腺过氧化物酶抗体（TPO-Ab）在甲状腺功能亢进（甲亢）、甲状腺功能减退（甲减）、单纯性甲状腺肿大患者的临床价值。方法：采用放射免疫分析检测甲状腺疾病患者血清TGA、TMA、TPO-Ab浓度及血清FL、FT4、TSH水平,分析TPO-Ab在69例甲亢、53例甲减及45例单纯性甲状腺肿的阳性率。结果：TPO-Ab的阳性率（82％-92．5％）明显高于同组的TGA（44．2％）、TMA（69．8％）,甲减组TPO-Ab的阳性值（平均57．4％）明显高于甲亢组（31．2％）和单纯甲状腺肿组（17．6％）,差异有显著意义（P〈0．01）。结诊!TPO-Ab对自身角癌件甲状腺疾病的诊断、治疗和预后评估具右一定的临床价值．     

500例自身免疫病患者抗核抗体谱检测与临床血液学检测的对比性研究

目的:探讨在自身免疫性疾病中抗核抗体线性免疫印迹分析法(ANA-LIA)与临床血液学检测的相关性.方法:抽取患者的血液,常规进行临床血液学检测及ANA-LIA法检测.ANA-LIA法使用德国IMTEC公司IMTEC-Human GmbH抗核抗体谱试剂盒.结果:在500例样本中,分别视疾病不同而有(1~5)项不同抗体阳性的为80例(男23,女57).年龄(12~83)岁,平均52岁,其检测ANA-LIA阳性的同时,临床血液学检测指标中性粒细胞组升高的22例(27.5%);淋巴细胞组升高的10例(12.5%);C反应蛋白组升高的34例(42.5%);血沉组升高的25例(31.2%);类风湿因子组升高的13例(16.2%);白细胞组升高的9例(11.2%);无改变组19例(23.7%).结论:在80例ANA-LIA检测阳性标本的同时,有61例临床相关血液检查出现了不同变化,与自身免疫性疾病有关也可能与其他潜在疾病相关.因此,在自身免疫性疾病筛查中将临床常规的血液学检测与ANA-LIA联合起来可能更有利于疾病的诊断.     

Immunosensors for Biomarker Detection in Autoimmune Diseases

Archivum Immunologiae et Therapiae Experimentalis - Autoimmune diseases occur when the immune system generates proinflammatory molecules and autoantibodies that mistakenly attack their own body....     

Evaluation of Five Antibody Detection Tests for Diagnosis of Bovine Paratuberculosis

Five diagnostic tests based on enzyme-linked immunosorbent assay (ELISA) technology for bovine paratuberculosis were evaluated by using individual serum or milk samples from 359 dairy cattle in seven paratuberculosis-free herds and 2,094 dairy cattle in seven Mycobacterium paratuberculosis-infected dairy herds. Three independent laboratories using three different culture procedures completed fecal cultures for M. paratuberculosis on these cattle and found 417 cows to be shedding M. paratuberculosis in their feces. An animal that was fecal culture positive for M. paratuberculosis by any of the three laboratories was considered a confirmed case of infection. The specificity of three ELISAs (two on serum and one on milk) was ≥99.8%. The specificity of the remaining two ELISAs, both done on serum, was 94.9 and 84.7%. Four of the five ELISAs evaluated produced similar sensitivity in detecting fecal culture-positive cattle (27.8 to 28.9%). Serum ELISA “D” had the lowest specificity (84.7%) and the highest sensitivity (44.5%), but if the cutoff value defining a positive test was changed from 125 to 250% (of the positive control) the sensitivity and specificity, 31.8 and 97.5%, respectively, were comparable to those of the other four assays. If the case definition for M. paratuberculosis infection was based on the culture results of a single laboratory instead of the combined results of three laboratories, ELISA sensitivity estimates were 45.7 to 50.0%. With the exception of ELISA D, assay agreement was high (kappa 0.66 to 0.85) for categorical assay interpretations (positive or negative), but linear regression of quantitative results showed low correlation coefficients (r² = 0.40 to 0.68) due to the fact that ELISA results for some cows were high in one assay but low in another assay. Likelihood ratio analysis showed a direct relationship between the magnitude of ELISA result and the odds of a cow shedding M. paratuberculosis in its feces. If used judiciously and interpreted quantitatively, these ELISAs are useful tools in support of paratuberculosis control programs in dairy herds.     

酶联免疫吸附试验检测梅毒螺旋体抗体的分析性能评价

目的评价ELISA检测梅毒螺旋体（TP）抗体的分析性能,探讨未知诊断的定性实验的性能评价方法。方法参照临床实验室标准化协会（CLSI）发布的EP12-A2文件对ELISA法测定TP的临界值±20%范围的重复性进行分析,并与TP明胶凝集试验（TPPA）结果进行一致性比较。结果 ＋20%浓度临界值检测阳性率为≥95%,CV为14.8%;-20%浓度临界值检测阴性率≥95%,CV为19.6%;两种方法的一致程度百分比为99.6%,一致程度95%可信区间为98.8%~99.6%。结论 TP-ELISA法检测TP抗体的临界值±20%浓度范围之外标本,可得到可靠的检测结果。TP-ELISA法与TPPA法结果一致性好,可代替其作为TP诊断试验。