979例自身免疫性疾病抗核抗体与抗ENA抗体联合检测的结果分析

目的回顾本实验室开展抗核抗体(antinuclear antibody,ANA)与抗可提取性核抗原(extractable nuclear antigens,ENA)抗体联合检测的情况,分析两者之间的相关性,并探讨联合检测在自身免疫性疾病中的应用价值。方法间接免疫荧光法检测ANA,生物芯片技术检测抗ENA(ds-DNA、Sm、SS-A/Ro、SS-B/La、u1RNP、Scl-70、Jo-1、RibosomalP)抗体,并采用双盲法分析ANA阳性标本的荧光核型与抗ENA抗体的关系。结果979例临床标本中,ANA阳性的标本数为400例,阳性率为40.9%(400/979)。荧光核型主要是核颗粒型(249例)、核均质型(83例)、胞浆颗粒型(30例),同时对979例标本进行抗ENA抗体的检测,阳性标本数为199例,阳性率为20.3%(199/979),只有1例阳性出现在ANA阴性的标本中。在400例ANA阳性的标本中,抗ENA抗体的阳性率为49.5%(198/400),出现较多的阳性抗体是抗SS-A/Ro(133例)、抗RibP(78例)、抗u1RNP(70例)、抗ds-DNA(61例)、抗SS-B/La(40例)等抗体,主要与颗粒型荧光相关。而均质型ANA标本中,常见的抗ENA抗体是抗ds-DNA(37例)、抗SS-A/Ro(27例)、抗Jo-1(20例)、抗RibP(20例)等抗体。结论不同荧光核型的ANA与抗ENA抗体之间的关系有各自的特点。用间接荧光法筛查ANA,并联合抗ENA抗体谱进行分型,可以确定部分ANA靶抗原的类型,进一步扩展抗ENA抗体谱,才能准确确定靶抗原。     

抗核抗体核型与抗核抗体谱检测结果对比分析

目的：分析本实验室抗核抗体（ANA）与抗核抗体谱（ANAs）17项联合检测的情况,并探讨两者之间的相关性。方法分别用间接免疫荧光法和免疫印迹法检测患者 ANA 和 ANAs,判断其 ANA 核型和 ANAs 结果,并对各荧光核型和各种特异性抗核抗体结果进行统计分析。结果340例 ANA 阳性患者标本中,颗粒型214例,着丝点型51例,胞浆颗粒型28例,核仁型25例,均质型17例,周边型5例,颗粒型是检出的主要核型。ANAs 17项抗体阳性率为82．94％。214例荧光颗粒型标本中,常见的抗体为抗 SSA/Ro52kd（31．78％）、抗 SSA/Ro60k（d30．84％）和抗 SSB/La（19．16％）抗体;17例荧光均质型标本中,常见的抗体为抗核小体（47．06％）和抗组蛋白（41．18％）抗体;抗 CenpB 对着丝点核型的诊断具有较高的特异性,阳性率为92．16％（47/51）。结论不同荧光核型的 ANA 与 ANAs 抗体之间有一定的关系,但并无绝对的规律可寻,两种 ANA 的检测方法,间接免疫荧光法和免疫印迹法联合应用,有助于自身免疫性疾病（AID）的诊断和预后观察。     

2513例抗核抗体与抗核抗体谱检测结果对比分析

目的分析本实验室抗核抗体(ANA)与抗核抗体谱(ANAs)15项联合检测的结果,并探讨两者之间的关系。方法分别用间接免疫荧光法(IIF)和免疫印迹法(IBT)检测患者ANA和ANAs,判断其ANA滴度、荧光模型和ANAs结果,并对ANA和ANAs检测结果进行对比分析。结果 471例ANA阳性患者标本中,颗粒型161例,均质型110例,胞质型96例、核仁型37例、着丝点型28例,其他型合计39例;滴度为1∶100者196例,占41.61%(196/471),其中ANAs阳性53例,占27.04%(53/196);滴度为1∶320者123例,占26.11%(123/471),其中ANAs阳性67例,占54.47%(67/123);滴度为1∶1 000者152例,占32.27%(152/471),其中ANAs阳性132例,占86.84%(132/152);2 042例ANA阴性标本中45例ANAs检测阳性,阳性率为2.2%(45/2042);高滴度ANA患者血清中ANAs的阳性率高于低滴度(χ2=123.132,P0.05)。结论不同滴度的ANA与ANAs之间有一定的关系,如果以ANA进行过筛,阳性再做ANAs会漏检部分患者,建议IIF和IBT联合应用,防止风湿病的漏诊。     

间接免疫荧光法与化学发光法检测抗核抗体的对比分析

目的 探讨间接免疫荧光法(IIFA)和化学发光法(CMIA)检测抗核抗体(ANA)的异同.方法 确诊为自身免疫性疾病患者血清240例,同时采用IIFA法和CMIA法检测ANA.对两种方法检测结果不符者采用欧蒙印迹法复查抗核抗体谱.结果 IIFA法与CMIA法检测240例ANA阳性率分别为90.83%和78.75%,差异有统计学意义(P<0.05);两种检测方法的符合率为77.92%;在53例两种方法检测结果不一致的标本中,41例IIFA(+)CMIA(-)患者抗核抗体谱阳性率仅为14.63%;而12例IIFA(-)CMIA(+)患者抗核抗体谱阳性率高达66.67%,差异有统计学意义(P<0.05);IIFA不同荧光核型中,"其他型" 在两种方法检测符合率方面低于颗粒型、胞浆型、均质型核型(P<0.05).结论 CMIA法检测总ANA的敏感度低于IIFA法,而CMIA法的特异度优于IIFA法,因此将IIFA法作为ANA的筛查方法,再结合CMIA法检测,可在获得荧光核型信息观察抗体滴度的同时,进一步对ANA进行定量.而且应用两种不同方法学原理的实验互相佐证,还可以减少实验误差,提高检测的准确度.     

间接免疫荧光法检测抗核抗体罕见核型的临床研究进展

摘要:抗核抗体(ANA)是自身免疫病重要的辅助诊断指标,间接免疫荧光法检测ANA仍被认为不可替代的经典方法。ANA荧光模型国际共识(ICAP)将间接免疫荧光法检测ANA细分为30种,其中一些常见的核型已被广泛研究,而罕见核型的临床意义和靶抗原虽有研究,但多为零星的病例报道,仍需要进一步综合评估。该文就罕见抗核抗体的核型特征、临床相关性及靶抗原进行综述,为罕见核型的临床应用提供依据。     

间接免疫荧光法与印迹法检测抗核抗体的相关性研究

目的研究间接免疫荧光法(IIF)检测抗核抗体(ANA)与印迹法(LIA)检测ANA谱(ANAS)的诊断结果,并分析二者的一致性及相关性。方法选取2013年2月至2014年11月在该院风湿科疑为自身免疫性疾病(AID)的住院及门诊患者为研究对象,采用IIF与LIA分别检测ANA与抗核抗体谱ANAS,并对结果进行统计分析。结果检出ANA阳性共127例,ANAS阳性84例,最终确诊AID 96例,各种特异性抗体均有检出,优势性检验发现总体阳性率差异有统计学意义。结论使用IIF检测ANA的阳性率高于LIA,但对于疾病的诊断价值,则LIA检测ANAS优于IIF检测ANA,临床上二者不可互相代替,应联合检测,防止漏检。     

血清抗核抗体荧光核型及抗核抗体谱检测在系统性红斑狼疮诊断中的应用价值分析

目的　研究抗核抗体（antinuclear antibody,ANA）荧光核型和抗体谱联合检测在系统性红斑狼疮（systemiclupus erythematosus,SLE）患者诊断中的应用价值。方法　对135 例SLE 患者、96 例非SLE 风湿性疾病患者与96 例健康体检者分别采用间接免疫荧光法（indirect immunofluorometric assay,IIF）和线性免疫印迹法（linearityimmunoblotting assay,LIA）检测血清中的ANA荧光核型和ANA谱,分析两种方法联合检测在SLE患者诊断中的应用价值。结果　SLE 病人ANA阳性率为99.3%,显著高于疾病对照组（75.0%）和健康对照组（4.5%）,差异有统计学意义（P<0.05）。ANA 荧光核型以核颗粒型为主（46.7%）,其次是核均质型（22.2%）,还有少量的胞浆型（13.3%）和均质胞浆混合型（11.1%）。ANA 谱分析发现双链DNA（double-strand DNA,dsDNA）抗体、抗低分子量核糖核蛋白（low-molecularribose nuclear protein,nRNP）抗体、抗核小体（nucleosome,NUC）抗体、Ro-52 抗体、SSA 抗体、抗核糖体P 蛋白（ribosme P protein, RIB）抗体、抗组蛋白（histone, HI）抗体、Smith（Sm）抗体和SSB 抗体九种抗体的敏感度均显著高于疾病对照组和健康对照组,两两比较差异均有统计学意义（χ2=6.60~83.74,均P<0.01）。其中dsDNA 抗体的敏感度和特异度分别为57.8% 和99.0%,nRNP 抗体的敏感度和特异度分别为48.9% 和95.8%,NUC 抗体的敏感度和特异度分别为40% 和99.0%,这三种抗体的敏感度和特异度都比较高。结论　① dsDNA 抗体、nRNP 抗体和NUC 抗体可以作为SLE 诊断的标志性抗体。② ANA 荧光核型分析敏感度高而特异度低,ANA 谱检测敏感度稍低而特异度高。这两种方法各有其优缺点,在临床工作中应把二者结合起来应用以利于SLE 的诊断和治疗。     

125例SLE患者血清抗核抗体及抗核抗体谱检测结果分析

目的探讨抗核抗体（ANA）及抗核抗体谱对系统性红斑狼疮（sLE）的临床价值。方法对1525例确诊的SLE患者血清ANA及ANA谱进行检测分析。ANA和抗双链DNA抗体采用间接免疫荧光法（IIF）,ANA谱采用欧蒙印迹法。结果①125例SLE患者血清ANA阳性共121例,阳性率为96．8％。②患者血清ANA谱大多为抗体二联及以上共同出现阳性,各抗体阳性率分别为抗一ul—nRNP／Sm：48．8％、抗一Smtl5．2％、抗一SS—A：36％、抗一R0—52：54．4％、抗一SS—B：14．4％、抗一scl—70：O．8％、抗一dsDNA：55．2％、抗一Neukleosmone：39．2％、抗一Histone：37．6％、抗一Rib—Pro!：12％、抗一Jo一1和抗一CENP—B未检出。结论ANA和ANA谱联合检测在SLE诊断中具有互补性,可提高检出率,对SLE的诊断分型、治疗、预后判断和病情随访等均有重要的临床价值。     

间接免疫荧光法检测抗核抗体与免疫印迹法检测抗核抗体谱结果的分析比较

目的比较间接免疫荧光法(IIF)筛查抗核抗体(ANA)与免疫印迹法(LIA)检测抗核抗体谱(ANA谱)特异性抗体结果的一致性,分析二者的相互关系及临床意义。方法2012年6月~2012年12月期间在中国中医科学院西苑医院风湿病科住院的503例患者,分析其ANA与ANA谱检测的一致性。将ANA与ANA谱的联合检测结果分为ANA+/ANA谱+、ANA+/ANA谱-、ANA-/ANA谱+、ANA-/ANA谱-四组,比较各组间的差异,并分析各类型自身免疫性疾病(AID)患者的分布情况。最后探讨AID患者ANA与ANA谱非一致时所检测出的核型及特异性抗体的特点。结果ANA与ANA谱检测结果无显著统计学差异,两者符合率为77.27%,不符合率为22.73%。ANA+/ANA谱+,ANA+/ANA谱-,ANA-/ANA谱+三组统计显示AID与非AID患者的概率不相同;ANA-/ANA谱-组AID与非AID患者的概率相同。结论ANA与ANA谱联合检测比单一检测时阳性预测值(PPV)提高,假阴性率(FNR)减低。ANA+/ANA谱+,ANA+/ANA谱-,ANA-/NA谱+三组对AID疾病有鉴别意义,而ANA-/ANA谱-组对AID无鉴别意义。     

间接免疫荧光法及免疫印迹法检测抗核抗体在SLE中的临床价值

目的研究间接免疫荧光法(IIF)筛查的抗核抗体(ANA)和免疫印迹法(IBT)检测的特异性ANA谱在系统性红斑狼疮(SLE)患者的相关性和临床意义。方法对80例SLE患者(SLE组)、80例其他风湿病患者(疾病对照组)及80例健康体检者(健康对照组)分别采用IIF、IBT检测血清中的ANA。结果 SLE组ANA阳性率(96.25%)显著高于疾病对照组(32.50%)和健康对照组(5.00%),差异有统计学意义(P0.05);SLE组荧光模式主要是颗粒型(43例,53.75%)、均质型(16例,20.00%)、胞浆颗粒型(14例,17.50%)。SLE组ANA的阳性率为97.5%,其中抗nRNP/Sm抗体、抗Sm抗体、抗dsDNA抗体、抗核小体抗体、抗组蛋白抗体、抗Rib-P抗体阳性率均显著高于疾病对照组和健康对照组,差异有统计学意义(P0.05)。结论 ANA和ANA谱联合检测对SLE的诊断及治疗具有重要意义。     

抗核抗体核型与条带免疫抗体谱相关性分析

目的分析抗核抗体(antinuclear antibody,ANA)与抗核抗体谱(antinuclear antibodies,ANAs)的相关性。方法 2 500例疑似或已确诊自身免疫性风湿性疾病患者,采用间接免疫荧光法检测ANA,其中826例同时应用条带免疫分析法行ANAs分析。结果 ANA阳性1 300例,同时行ANAs的826例患者中ANA(+)/ANAs(+)182例(22.03%),ANA(-)/ANAs(-)529例(64.04%),ANA(+)/ANAs(-)111例(13.43%),ANA(-)/ANAs(+)4例(0.50%);ANA阳性者中以颗粒型(62.2%)、胞质颗粒型(13.3%)、均质型(9.7%)、核仁型(8.9%)多见,颗粒型多见SSA/Ro52抗体,均质型多见dsDNA抗体,胞质颗粒型多见AMA-M2;ANAs的阳性率随ANA滴度增大而升高;当滴度为1∶800以上时,>20～40岁组ANA阳性率明显高于其他各年龄组(P<0.01)。结论 ANA-间接免疫荧光法是良好的筛查实验,ANA核型、滴度与ANAs相结合对自身免疫性风湿性疾病的诊断与鉴别诊断有重要意义。     

Clinical significance of anti-neutrophil cytoplasm antibodies detected by a standardized indirect immunofluorescence assay

We assessed the use of a standardized anti-neutrophil cytoplasmantibody (ANCA) test in diagnosing Wegener's granulomatosis(WG), microscopic polyarteritis (mPA) and systemic vasculitis(SV). All samples (n = 779) tested for ANCA at our laboratorywere identified, and clinical information was obtained for 783/779patients by questionnaire, and by visits where necessary. Thecombined prevalence of WG/mPA/SV was 123/738 (17%). The ANCAtest was positive in 48/68 WG patients (71 %; 38 cANCA, 10 pANCA),22/43 mPA patients (51%; 12 cANCA, 10 pANCA) and 3/12 SV patients(25%). WG and mPA patients in remission had similar frequenciesof positive ANCA to those with active disease. The sensitivityand specificity for WG (71 % and 80%) and mPA (51% and 80%)were lower than previously reported. In this high-prevalencepopulation, the overall (WG/mPA/SV) positive predictive valuewas only 40%, and the sensitivity 59%. Only 29% of positivetests were from patients with active disease. Overall, 78% oftest results gave a ‘true’ prediction. On this basis,a diagnosis of necrot-izing vasculitis (WG/mPA/SV) can be neithermade nor refuted by ANCA test alone.     

1801例抗核抗体检测结果及其临床分析

目的 探讨抗核抗体(ANA)的流行病学特征及其与临床的相关性.方法 回顾性分析1 801例ANA检测结果及其临床资料.结果 (1)ANA阳性604例,总阳性率为33.5%,其中82.5%为女性患者.60岁以下各年龄组女性患者阳性率均高于男性(P<0.05),而以36～45岁组阳性率最高.(2)ANA荧光模型以细胞核型荧光(67.4%)最常见,以核颗粒型(43.7%)为主.(3)ANA阳性患者特异性抗体总阳性率为47.8%.混合型特异性抗体阳性率最高(57.8%),细胞浆型(12.7%)最低(P<0.01).抗核点型、抗核膜型、抗着丝点型及抗中心粒抗体阳性患者均未检出特异性抗体.(4)ANA阳性者66.2%诊断为自身免疫性疾病(AID),显著高于ANA阴性者(17.0%),差异有统计学意义(P<0.01).混合型ANA组73.6%诊断为AID,其次为细胞核型ANA组(70.2%),最低为细胞浆型ANA组(36.9%),后者与前两组比较,差异有统计学意义(P<0.01).系统性红斑狼疮(SLE)组ANA阳性率最高.SLE、类风湿关节炎(RA)、混合结缔组织病(MCTD)、系统性硬皮病(SS)、多发性肌炎(PM)/皮肌炎(DM)、重叠组织病患者ANA阳性率与非AID患者比较,差异有统计学意义(P<0.05),而强直性脊柱炎(AS)、成人斯蒂尔病(STILL)患者与非AID患者比较,差异无统计学意义.SLE、RA、MCTD患者ANA模型均以核颗粒型为主.滴度达1∶3 200者89.1%诊断为AID,其次为滴度达1∶1 000者(72.2%)与滴度达1∶320者(44.7%),3组间比较,差异均有统计学意义(P=0.000).结论 不同性别和年龄患者ANA阳性率存在差异;ANA存在多种荧光模型,不同荧光模型ANA阳性患者特异性抗体检出率存在差异,部分ANA阳性患者不能检测出特异性抗体;高滴度ANA对于AID诊断有重要意义,但ANA荧光模型对不同AID疾病的诊断无特异性.     

Detection of anti-SSA antibodies by indirect immunofluorescence

BACKGROUND: HEp-2 cells that overexpress the human 60-kDa SSA antigen have been used to improve sensitivity and specificity for the detection of anti-SSA antibodies by indirect immunofluorescence. We describe a survey on the detection of anti-SSA antibodies using a commercial substrate that overexpresses SSA. METHODS: The evaluation was done on 18 371 consecutive samples submitted to the laboratory for detection of anti-nuclear antibodies, from which 188 anti-SSA antibody-containing and clinically documented samples were obtained. The presence of anti-SSA antibodies produced a distinct bright speckled pattern with nucleolar staining in 10-20% of interphase cells. The identity of all anti-SSA antibodies was confirmed by dot-blot analysis. RESULTS: Samples containing anti-SSA antibodies were separated into three main groups: group I, distinctive SSA pattern and other nuclear staining (50%); group II, only the distinctive SSA pattern (29%); group III, nuclear staining but without the distinctive SSA pattern (21%). Anti-SSA antibodies with concurrent SSB antibodies were associated with group I, whereas anti-SSA antibodies with concurrent U(1)-RNP antibodies were associated with group III. Group I included mainly patients with Sjogren syndrome and systemic lupus erythematosus (SLE), whereas group III included patients with mixed connective tissue disease and SLE. Diseases not classically associated with the presence of anti-SSA antibodies were found in group II in >50% of the cases. CONCLUSIONS: SSA-positive individuals were identified in a population selected on the basis of HEp-2000 positivity. Our study highlights diseases associated with anti-SSA antibodies and associations between the presence of the distinctive SSA pattern on HEp-2000 and some clinical conditions.     

Comparison of antinuclear antibody profiles obtained using line immunoassay and fluorescence enzyme immunoassay

ObjectiveLIA-ANA-Profile-17S is a multiplex line immunoassay that simultaneously detects 17 antinuclear antibodies (ANAs) against extractable nuclear antigens (ENAs). We evaluated the utility of LIA-ANA-Profile-17S as a supplement to ANA indirect immunofluorescence (IIF) and EliA ENA (a fluorescence enzyme immunoassay) for diagnosis of ANA-associated rheumatic diseases.MethodsSera were collected from 245 patients referred for an ANA IIF test. LIA-ANA-Profile-17S results were compared with those of EliA ENA. The kappa coefficients, agreement rates, and diagnostic performance of these tests were assessed for systemic lupus erythematosus (SLE) and Sjögren’s syndrome (SjS).ResultsWe observed almost perfect interassay agreement for antibodies against Ro52/Ro60, CENP-B, and Scl-70 (kappa = 0.91, 0.97, and 1.00, respectively); strong agreement for anti-SS-B/La antibody (kappa = 0.81); and relatively low agreement for other antibodies, including those against dsDNA, Sm, RNP, and Jo-1. For SLE diagnosis, LIA-ANA-Profile-17S showed lower sensitivity and similar specificity compared with EliA ENA. The sensitivity and specificity of these two assays were similar for SjS diagnosis.ConclusionsThe specificity of LIA-ANA-Profile-17S was enhanced when combined with ANA IIF and was comparable with that of EliA ENA. LIA-ANA-Profile-17S showed relatively good agreement with EliA ENA. In combination with ANA IIF, these assays showed enhanced diagnostic performance.     

Detection of human anti-HTLV-III antibodies by indirect immunofluorescence using fixed cells

Seropositivity to human T-cell lymphotrophic virus-III (HTLV-III) can have profound implications for the individual in whom it is detected. Simple and reliable tests are needed to confirm positivity by screening assays. In this study, detection of human antibodies by indirect immunofluorescence (IFA) on acetone-fixed HTLV-III infected H9 cells was evaluated in blood donors, patients with infectious or auto-immune diseases, and men with or at high risk for developing acquired immune deficiency syndrome (AIDS). Specific and nonspecific patterns of immunofluorescent reactivity were distinguished easily. None of 98 serums from blood donors was positive, while two of 33 serums from patients attending an infectious disease clinic, both homosexual men, were positive. Ninety-six percent of 24 serums from men with AIDS, 87 percent of 70 serums from men with lymphadenopathy, and 50 percent of 135 serums from healthy gay men were positive. These results paralleled those obtained by Western blotting and membrane immunofluorescence. In contrast, 11 and 4 percent, respectively, of these serums were judged as borderline or not interpretable by an enzyme-linked immunosorbent assay (ELISA). Of these serums, those that were positive by IFA were positive by Western blots, and 16 of the 17 IFA-negative serums were negative by Western blots. These studies indicate that IFA is a sensitive and specific assay for HTLV-III antibodies in human serums.     

Comparative study of results of serological diagnosis of Lyme borreliosis by indirect immunofluorescence and immunoenzyme analysis

A total of 176 sera from 73 patients with verified Lyme borreliosis at different stages of the disease are examined. Serological diagnosis was carried out by 2 methods: indirect immunofluorescence (IIF) with corpuscular B. burgdorferi antigen and enzyme immunoassay (EIA) with purified flagellar B. burgdorferi antigen (Dako). EIA with Dako antigen is more sensitive for the diagnosis of Lyme borreliosis at any period of the disease than IIF. Analysis of correlations between the results of IIF and EIA showed correlation in the levels of IgG but not IgM antibodies. The findings confirmed a previous hypothesis that inadequate antibacterial therapy before investigation decreases the level of antibodies to Borrelia. In patients with a history of Lyme borreliosis, antibodies to B. burgdorferi are detected less frequently by both IIF and EIA. Patients with persistent levels of antibodies to B. burgdorferi, even without clinical signs of infection, are in need of regular check-ups, because the prognostic significance of antibodies to B. burgdorferi is unknown and relapses may occur after months and years.