抗卵细胞核抗体的研究

系统性自身免疫疾病患者体内出现的抗核抗体可与多种体细胞折胞核发生反应已有较多研究，但其是否可以与卵细胞核发生反应的研究尚少见报道。以鸡卵核为抗原建立了酶联免疫法，检验相应抗体。结果表明；体细胞ＡＮＡ阳性组卵核ＡＮＡ的阳性率为８３．３％，体细胞ＡＮＡ阴性组卵核ＡＮＡ的阳性率为２３．３％，两者具有显著性差异，从而证实ＡＮＡ亦可与卵细胞胞核发生反应。     

斑点金免疫渗滤法检测血清抗精子抗体

目的建立检测血清抗精子抗体的斑点金免疫渗滤法 (dot im munogold filtration assay,DIGFA)。方法　采用经 Sephadex G- 75 -抗人全血清柱亲和层析纯化的人精子抗原和胶体金标记的 SPA,根据免疫渗滤原理 ,建立抗精子抗体(ASAb)的斑点金免疫渗滤法检测法。结果　纯化的人精子抗原具有较好的特异性 ;斑点金免疫渗滤法与 EL ISA法比较 ,符合率为 97.7%。特异性为 96 .2 % (TN/ TN+ FP× 10 0 % ) ,敏感性为 88.9% (TP/ TP+ FN× 10 0 % ) ,阳性预测值 84.2 % (TP/TP+ FP) ,阴性预测值 97.2 % (TN/ TN+ FN) .结论　斑点金免疫渗滤法方法检测血清抗精子抗体有较好的特异性和敏感性 ,重复性好 ,试剂稳定 ,操作简便快速 ,具有临床应用价值     

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。     

抗核抗体核型分布及核型与特异性抗体相关性分析

目的通过对ANA核型分布及核型与特异性抗体相关性分析,为临床对ANA和ANA谱检测结果判读提供参考。方法用间接免疫荧光法检测ANA,免疫印迹法检测ANA谱,对449例ANA或ANA谱阳性的结果进行核型和特异性抗体分析。结果 ANA核型分布以核颗粒型为主,占78.2%;核型与特异性抗体有一定的相关性,但同一种自身抗体可出现不同的荧光核型。结论 ANA和ANA谱同时检测能提高AID的检出率,有助于准确的判断特异性抗体,对AID的诊断、病情监测、预后判断有十分重要的作用。     

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.     

用免疫金银法检测抗核抗体

抗核抗体(ANA)是诊断风湿性疾病的常用实验室指标。经典的检测方法为间接荧光法(IFA)~。但此法受荧光显微镜、荧光抗体、底物(抗原)片的种类与质量影响甚大,荧光又极易淬灭,标本片不能长期保存。我们建立了一种免疫金银法(IGSS)测定ANA,操作简便,结果清晰,无需特殊设备,一般实验室均可采用。     

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。     

斑点免疫金银染色检测抗双链DNA抗体

从牛胸腺ＤＮＡ制备双链ＤＮＡ溶液，点于混合纤维素酯微孔滤膜，进行斑点免疫金银染色（Ｄｏｔ－ＩＧＳＳ），以检测抗双链ＤＮＡ抗体（ＡｄｓＤＮＡ）。在３１６份所测血清中此法检出的ＡｄｓＤＮＡ对ＳＬＥ的敏感性、特异性分别是６９．４％、９６．８％，与平行对照的ＥＳＮ／ＡＥＳ－ＩＧＳＳ、ＲＩＡ、ＥＬＩＳＡ等法相比差异无显著性（Ｐ＞０．０５），符合率均＞９２％。此法操作方便、结果观察简单明了，更为实用。     

抗核抗体谱的检测在临床应用中的研究进展

自身免疫性疾病（autoimmune diseases,AID）确切的发病机制目前尚不清楚,且临床症状和体征不典型,对其诊断和鉴别诊断较复杂,但由于该类疾病都存在一些特异性的自身抗体,所以对自身免疫性疾病的诊断不仅依赖于临床表现,而且很大程度上取决于免疫学实验诊断,特别是实验室的抗核抗体（antinuclear antibody,ANA）检测[1].通过对ANA类型及其滴度的检测,能为自身免疫性疾病的诊断、分型、治疗和预后判断提供重要的依据,本文就ANA和ANA谱的检测方法及其临床应用做一综述.     

全人单克隆抗核抗体的制备和鉴定

目的 本研究旨在从系统性红斑狼疮(systemmic lupus erythematosus,SLE)患者外周血淋巴细胞中获得自身抗体,并从中鉴定筛选出抗核抗体.方法 利用人B细胞杂交瘤技术制备SLE患者来源的全人单克隆自身抗体,采用酶联免疫吸附实验(enzyme-linked immuno sorbenk assay,ELISA)方法测定阳性杂交瘤细胞上清抗体浓度,并以HEp-2细胞为底物,采用间接免疫荧光法鉴定筛选出抗核抗体.结果 共获得48株稳定分泌IgG型全人单克隆自身抗体的杂交瘤细胞株,其中12种细胞上清抗体浓度大于1 500 ng/mL,占25％,通过免疫荧光鉴定从中筛选出3种抗核抗体,荧光核型分别为周边型、斑点型和核仁型.结论 全人单克隆自身抗体的制备及抗核抗体的筛选鉴定,为开发新型单克隆抗体药物以及研究SLE的致病机制奠定了良好的实验基础.     

检测抗核抗体对系统性红斑狼疮的临床应用

目的探讨检测抗核抗体对系统性红斑狼疮的临床价值。方法采用回顾性分析的方法,分析我院收治的系统性红斑狼疮患者的临床资料。结果 SLE患者核颗粒型、核均质型、浆颗粒型、浆均质型、核仁型、着丝点型各型阳性率均高于对照组,SLE组总阳性率明显高于对照组,P<0.05。两种狼疮治疗后ANA含量均较治疗前有所降低,但神经性狼疮ANA降低效果明显优于狼疮肾炎患者,P<0.05,差异均有统计学意义。结论 ANA可以作为系统性红斑狼疮患者发生、发展的临床预警指标,值得临床推广检测。     

抗核抗体系列检测对狼疮性肾炎进行鉴别诊断的临床意义

目的探讨抗核抗体(ANA)系列指标在狼疮性肾炎(LN)患者中的表达情况及临床意义.方法对406例系统性红斑狼疮(SLE)患者(其中LN 122例)和74例其他自身免疫病患者及120例健康体检者采用间接免疫荧光法测定ANA,应用欧蒙印迹法测定ANA系列.结果 SLE患者ANA阳性率为94.49%,其他自身免疫病组ANA阳...     

Metabolic mediators determine the association of antinuclear antibody subtypes with specific clinical symptoms in systemic sclerosis

PurposeThe aim of this study was to investigate the possible link between different types of systemic sclerosis-specific antinuclear antibodies, adipokines and endothelial molecules which were recently found to have a pathogenic significance in systemic sclerosis.Materials/methodsSerum concentration of adiponectin, resistin, leptin, endothelin-1, fractalkine and galectin-3 were determined in the sera of patients with systemic sclerosis (n ​= ​100) and healthy controls (n ​= ​20) using ELISA.ResultsThe following associations between antinuclear antibodies and increased serum concentrations were identified: anticentromere antibodies with endothelin-1 (p ​< ​0.0001; mean level in patients 2.21 vs control group 1.31 ​pg/ml), anti-topoisomerase I antibodies with fractalkine (p ​< ​0.0001; 3.68 vs 1.68 ​ng/ml) and galectin-3 (p ​= ​0.0010, 6.39 vs 3.26 ​ng/ml). Anti-RNA polymerase III antibodies were associated with increased resistin (p ​< ​0.0001; 15.13 vs 8.54 ​ng/ml) and decreased adiponectin (p ​< ​0.0001; 2894 vs 8847 ​ng/ml).ConclusionIn systemic sclerosis metabolic and vascular factors may serve as mediators between immunological abnormalities and non-immune driven clinical symptoms.     

组蛋白诱导抗核抗体产生及其对肾脏损害

目的 寻找系统性红斑狼疮中诱导抗核抗体（ANA）生成的真正的自身核成分免疫原。方法 从Con A活化的脾淋巴细胞中提取组蛋白免疫同系BALB／c小鼠,ELISA方法测定IgG类抗组蛋白、抗dsDNA抗体,免疫荧光法检测抗核抗体核型和免疫复合物在肾小球内的沉积,免疫印迹法测定抗可溶性核抗原抗体,考马斯亮蓝法检测尿蛋白含量,光镜下观察肾脏形态变化,电镜下观察肾小球电子致密物。结果 活性组蛋白诱导IgG类抗组蛋白、抗dsDNA等多种抗核抗体生成,且诱发同系小鼠产生SLE样肾脏病理变化。结论 活性组蛋白是诱发抗核抗体生成、造成肾损害的免疫原之一。     

抗核抗体系列及补体检测对狼疮性肾炎的诊断意义

目的 探讨抗核抗体系列及补体检测对系统性红斑狼疮(SLE)伴狼疮性肾炎(LN)诊断的意义.方法 对1 699例SLE患者和120例健康体检者采用间接免疫荧光法测定抗核抗体(ANA),应用欧蒙印迹法测定抗核抗体系列,应用散射比浊法测定补体C3、C4.结果 1699例SLE患者,LN组921例,ANA阳性率为97.4％;不伴LN组778例,ANA阳性率为98.2％;与对照组比较,r=0.983,P=0.001,差异有统计学意义.LN组的抗组蛋白抗体,抗核小体抗体,抗双链DNA抗体阳性率分别为49.3％、59.8％、63.3％;不伴LN组分别为21.5％、35％、47.9％;差异有统计学意义(r=0.452,P=0.007).抗线粒体-2抗体和抗着丝点抗体阳性率,LN组与不伴肾炎组比较差异有统计学意义(r=0.291,P=0.015).当抗组蛋白抗体+抗核小体抗体+抗双链DNA抗体同时阳性时,LN患者其补体C3,C4水平比这三种抗体非同时阳性时的水平均更低,差异有统计学意义(r=0.583,P=0.009).结论 抗核抗体系列及补体的血清学检测对诊断LN及其预后判断、疗效观察等具有重要意义.     

Evaluation of antinuclear antibodies by indirect immunofluorescence and line immunoassay methods′: four years′ data from Turkey

The presence of antinuclear antibodies (ANAs), directed against intracellular antigens, is a hallmark of systemic autoimmune rheumatic diseases. The indirect immunofluorescence (IIF) assay is among the most commonly used routine methods for ANA detection as the screening test. The objective of the study was to evaluate ANA patterns in a 4‐year period retrospectively. All 19 996 serum samples that were sent to the Laboratory of Medical Microbiology of the tertiary Hospital by any hospital department between 1 January 2009 and 1 January 2013 with a request to test for ANA, anti‐ENA or both were included in the study. Of these samples, 4375 (21.9%) were ANA‐IIF‐positive and 15621 (78.1%) were ANA‐IIF‐negative. The presented ANA‐positive samples consisted of 2392 (54.67%) homogenous, 818 (18.70%) speckled, 396 (9.05%) centromere, 242 (5.53%) nucleolar, 213 (4.87%) nuclear dots, 178 (4.07%) cytoplasmic (except for actin and golgi), 24 (0.55%) actin, 9 (0.21%) golgi, 53 (1.21%) nuclear membrane and 50 (1.14%) mixed pattern. Totally 7800 samples were examined by LIA. Of these samples, 3440 were positive and 4307 were negative with IIF and LIA. In addition, 22 samples were detected as IIF‐positive but LIA‐negative, whereas the rest 31 samples were IIF‐negative but LIA‐positive. ANA patterns in 22 IIF‐positive samples were homogenous (9), speckled (5), golgi (4), cytoplasmic (3) and nucleolar (1). SSA/Ro‐52, SSB/La and Scl‐70 positivity were detected in 31 IIF‐negative/LIA‐positive samples by LIA. The present study comes forward with its overall scope, which covers 4‐year data obtained in tertiary hospital located in the western part of Turkey.     

Antinuclear antibodies: two-step detection strategy

In order to evaluate the performance of the chemiluminescent immunoassay (CLIA) in antinuclear antibodies (ANA) testing, using indirect fluorescent antibody (IFA) on HEp-2 cells as a standard, 209 samples were studied from September to October/2010. The tests were conducted according to the procedures recommended by the manufacturers of the reagents. The interpretation of the IFA results was done according to the Brazilian standards. The charts of patients showing different results between the two techniques were analyzed. The CLIA efficiency was 89%, with a sensitivity of 65%, and a specificity of 94.7%. Nine (4.3%) false-positive and 14 (6.7%) false-negative results were detected. Of these, 13 (93%) represented no risk for the diagnosis and therapeutic management of the patients. The CLIA methodology reduced the need for the IFA manual technique by 77% (160/209). The ANA screening test proved to be a fast and acceptable methodology in the studied population. We established the following criteria for the introduction of an automated ANA screening: (1) Positive results must be confirmed by IFA to characterize the pattern and titer of the antibody. (2) Negative results are issued with a notice to request a new test by IFA when the clinical suspicion of autoimmune disease persists.