血清PSA、f-PSA及f-PSA/PSA CLIA测定对前列腺疾病的临床应用价值

前列腺特异抗原(prostate spec ific antigen,PSA)是与前列腺癌相关的一种抗原。在前列腺癌的早期诊断、疗效观察及预后判断上越来越受到临床医生的重视。本院自2003以来,以PSA、f-PSA及f-PSA/PSA诊断前列腺疾病132例,现报告如下。1材料和方法1·1材料1·1·1正常对照组56例,年     

电化学发光法测定血清CA125对卵巢恶性肿瘤的诊断价值

卵巢恶性肿瘤是妇科常见肿瘤 ,其死亡率在妇科恶性肿瘤中占首位[1 ] ,而且 ,近年来发病率有明显上升趋势[2 ] 。目前尚缺乏敏感及特异的实验诊断和监测方法。我们采用电化学发光法测定血清ＣＡ12 5含量 ,以探讨血清ＣＡ12 5对卵巢恶性肿瘤的诊断价值尤其早期诊断价值 ,现报道如下。对象和方法一、对象 :(一 )正常对照组 :5 0例 ,年龄 2 1～ 6 3岁 ,平均 4 2 6岁 ,均为我院正常健康体检职工。(二 )良性对照组 :86例 ,年龄 2 3～ 5 1岁 ,平均 4 0 5岁 ,均为我院住院病人 ,术后病理检查证实为良性肿瘤 ,其中子宫肌瘤 38例 ,卵巢巧克力囊肿 2 …     

电化学发光法检测血清β-HCG的临床应用评价

我院采用电化学发光免疫分析法(ECLIA)定量测定血清β-HCG,现对该法的临床应用评价如下。1资料和方法1·1对象从2004年3月~2005年2月在我院受检的患者,共100例,其中健康非孕妇女30例,早孕25例,宫外孕35例,葡萄胎6例,绒癌4例。年龄(21~41)岁,平均(29.41±5.27)岁。1·2方法取患者静脉血3m l,分离血清,用ECLIA完成检测,ECLIA应用Roche E lecsys2010检测仪,试剂由罗氏公司提供。2结果2·1线性试验将ECLIA法测定的β-HCG值在(1112.01~3474.46)m IU/m l范围内的8份血清混合,反复测定6次,取其均值作为该混合血清的理论值。将该混合血…     

血清CEA、TNF、TSGF联检在肺癌诊断的临床价值

肺癌是一种恶性程度高、发展迅速的临床常见肿瘤。临床上的早期诊断颇受科研工作者的关注。本文对肺癌患者进行放疗前后血清CEA、TNF、TSGF进行联检 ,以进一步探讨它在肺癌诊断和治疗中的临床价值。对象和方法一、对象 :(一 )正常人 :30人 (男 2 0 ,女 1 0 ) ,均为保健科体检合格的健康人 ,无心、肝、肺、肾等重要脏器疾患 ,肝肾功能试验正常。(二 )病人组 :36人。均为肿瘤科明确诊断的病人 ,最后全部均病理切片证实。其中鳞癌 1 6例 ,腺癌 1 2例 ,小细胞未分化癌 8例。分别于放疗前和放疗后 2个月空腹抽取静脉血3ml,离心分离血清后 ,-…     

电化学发光免疫分析法测定TSH、FT3、FT4的临床应用

电化学发光免疫分析法(electro-chemiluminescence immunoassay, ECLI)是继放射免疫、酶免疫、荧光免疫、化学发光免疫分析法之后的新一代标记免疫测定技术,是电化学发光(ECL)和免疫测定相结合的产物[1].它包括了电化学和化学发光两个过程.整个测定过程全自动控制,反应时间短,15～30分钟即可出结果.该法灵敏度高,检测下限可达1pmol/L.     

全自动电化学发光免疫分析法检测血清TRAb的方法学评价

目的:对电化学发光免疫分析(ECLIA)检测血清促甲状腺激素受体抗体(TRAb)进行方法学评价.方法:采用ECLIA分别检测62例GD患者、65例非GD患者以及41例健康对照者血清中TRAb的含量,评估ECLIA检测TRAb的精密度、回收率、诊断敏感性和特异性及与ELISA的相关性.结果:ECLIA检测TRAb的批内、...     

血清CA125在诊断卵巢癌、宫颈癌、乳腺癌的临床价值

CA125是近年研究较多的与卵巢癌的相关性肿瘤标志物,具有较高的阳性率,而其它肿瘤(乳腺癌、宫颈癌)有一定的阳性率,但其特异性、灵敏度均低于卵巢癌。本文应用CA125对卵巢癌、宫颈瘤、乳腺癌患者血清标本进行测定,现将结果报告如下。1材料和方法1·1对象选自云南省省肿瘤医院20     

血清SIgA放射免疫分析在肝病诊断中的价值

近几年有关SIgA在各种分泌液中含量的测定及其临床意义报道较多，但有关血清中SIgA测定的报道较少，本文对115例肝病患者及587例正常成年人用RIA测定血清中SIgA的含量，并就其临床价值进行探讨。     

血清CEA和CA125联检对卵巢癌诊断价值的探讨

血清肿瘤标志物CEA和CA12 5已广泛应用于临床 ,本文旨在探讨对 16 0例卵巢癌患者进行CEA、CA12 5联检在卵巢癌早期诊断及筛查的价值。1　材料和方法1 1　标本来源1 1 1　样本组　 16 0例卵巢癌患者均来自本院住院及门诊病人 ,年龄 (2 4～ 76 )岁 ,平均 5 0岁。1 1 2　对照组     

血浆CA125、SF和血清铁测定对卵巢肿瘤患者诊断的临床价值

近年来,本文探讨了血浆CA125、血清铁蛋白（SF）和血清铁（Fe）水平对卵巢肿瘤患者诊断的临床价值,现将结果报道如下。1材料和方法1.1一般资料选自我院门诊和住院的75例卵巢肿瘤患者,均经临床明确诊断（其中包括CT、MRI和病理学诊断证实）,其中卵巢癌患者36例,年龄（21～70）岁,平均（44.5±13.1）     

促甲状腺素光激化学发光免疫测定法的建立和初步应用

采用光激化学发光免疫测定法(LICA)技术建立促甲状腺素(TSH)快速定量检测方法。采用两株针对TSH不同表位的单克隆抗体,一株单抗包被发光微粒,另一株为生物素化单抗,两者与链亲和素包被的感光微粒一起构建双抗体夹心LICA。数据处理采用双对数函数处理程序。方法的灵敏度为0.015mIU/L;批内CV为2.5%～3.6%,批间CV为2.6%～4.4%;平均回收率为100.60%。与时间分辨荧光免疫分析法(TRFIA)比对,相关系数达0.9681;与TRFIA临床测定值呈明显相关;正常值范围为0.33～3.09mIU/L。本文建立的TSH光激化学发光免疫测定法是目前TSH检测中最快速灵敏的方法之一,该方法稳定性好,具有很好的应用前景。     

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.     

Suppression of autoimmune thyroid disease by FK 506: influence on thyroid-infiltrating cells,adhesion molecule expression and anti-thyroglobulin antibody production

Autoimmune thyroid disease was induced in female PVG/c rats by neonatal thymectomy, followed by sublelhal, whole body x-irradiation. Disease development, assessed by histological evidence of lymphocytic thyroiditis and circulating levels of anti-thyroglobulin antibodies, was reduced significantly by a 3-week course of FK 506 (0.5 or 1.5 mg/kg per day) commencing after the detection of autoantibody production. Thyroid-infiltrating mononuclcar cells (MNC) in untreated rats stained predominantly for CD4⁺ and MHC class II antigen which was expressed widely on dendritic cells. Fewer infiltrating cells expressed TCR α/β, CD5, CD8 or LFA- 1β. Intercellular adhesion molecule-1 (ICAM-1) was observed on MNC, vascular endothelial cells and a minoritiy of residual thyroid epithelial cells. FK 506 administration reduced markedly the incidence of infiltrating TCRα/β⁺, CD5⁺, CD4⁺, CD8⁺, and LFA-1β⁺ cells and the expression both of MHC class II antigens and ICAM-I on MNC, endothelial cells and thyrocytes. Compared with normal PVG/c rats, there were reduced incidences of CD4⁺ CD8^? and CD4^? CD8⁺ lymphocytes and an elevation in the CD4⁺/ CD8⁺ cell ratio in the spleens of animals with autoimmune thyroiditis. These changes were partially reversed by FK 506. Systemic drug levels estimated by enzyme immunosorbent assay were in excess of those known to blockade cytokine production by CD4⁺ T lymphocytes in vitro and some evidence of minor renal dysfunction was observed. The results are consistent with a therapeutic effect of FK 506 mediated via interference with CD4⁺ T lymphocyte function and adhesion molecule-dependent cytotoxic effector mechanisms.     

Substitution of antibody with molecularly imprinted 96-well plate in chemiluminescence enzyme immunoassay for the determination of chloramphenicol residues

A direct competitive chemiluminescence enzyme immunoassay was developed by using the molecularly imprinted 96-well plate as an artificial antibody to detect chloramphenicol (CAP). The artificial antibody was synthesised on the well surface of MaxiSorp polystyrene 96-well plate and CAP was conjugated with the horseradish peroxidase. The results showed that the imprinted plate exhibited antibody-like binding ability. The plate showed fast adsorption rate, 66% adsorption was finished within 20 min. The cross-reactivity for CAP, florfenicol and thiamphenicol were 100%, 1.25% and 2.08%, respectively. And the imprinted plate could be reused for many times without loss of sensitivity. The IC₅₀ and the detection limit values under optimum conditions were 30±2 µg·L^?1 and 0.9±0.01 µg·L^?1, respectively. The plate was used to detect CAP in sea cucumber, which showed excellent recoveries ranging from 89% to 98.7%. And the result correlated well with that obtained by the CAP enzyme-linked immunosorbent assay (ELISA) kit.     

Detection of H-2 and Sendai virus antigens by chemiluminescence

Two monoclonal antibodies reacting with Ia9 and H2.2, respectively, have been added to spleen cell suspensions prepared from mice of different H-2 haplotypes. Cellular light emission was monitored in a liquid scintillation spectrometer operated in the out-of-coincidence mode. The antibodies stimulated chemiluminescence (CL) in cells possessing the target antigenic determinant, but were inactive in cells lacking the determinant. In addition, CL could be stimulated in Sendai virus-infected spleen cells with anti-Sendai antibodies. The results demonstrate that CL measurement is a sensitive and simple method for the detection of cell surface antigens and for the screening of antibodies reacting with these antigens.     

Localization of key amino acid residues in the dominant conformational epitopes on thyroid peroxidase recognized by mouse monoclonal antibodies

Autoantibodies to thyroid peroxidase (TPO), the major target autoantigen in autoimmune thyroid diseases, recognize conformational epitopes limited to two immunodominant regions (IDRs) termed IDR-A and -B. The apparent restricted heterogeneity of TPO autoantibodies was discovered using TPO-specific mouse monoclonal antibodies (mAbs) and later confirmed by human recombinant Fabs. In earlier studies we identified key amino acids crucial for the interaction of human autoantibodies with TPO. Here we show the critical residues that participate in binding of five mAbs to the conformational epitopes on the TPO surface. Using ELISA we tested the reactivity of single and multiple TPO mutants expressed in CHO cells with a panel of mAbs specifically recognizing IDR-A (mAb 2 and 9) and IDR-B (mAb 15, 18, 64). We show that antibodies recognizing very similar regions on the TPO surface may interact with different sets of residues. We found that residues K713 and E716 contribute to the interaction between mAb 2 and TPO. The epitope for mAb 9 is critically dependent on residues R646 and E716. Moreover, we demonstrate that amino acids E604 and D630 are part of the functional epitope for mAb 15, and amino acids D624 and K627 for mAb 18. Finally, residues E604, D620, D624, K627, and D630 constitute the epitope for mAb 64. This is the first detailed study identifying the key resides for binding of mAbs 2, 9, 15, 18, and 64. Better understanding of those antibodies' specificity will be helpful in elucidating the properties of TPO as an antigen in autoimmune disorders.     

Comparison of chemiluminescence microparticle immunoassay,indirect immunofluorescence assay,linear immunoassay and multiple microbead immunoassay detecting autoantibodies in systemic lupus erythematosus

The aim of study was to detect antinuclear antibodies (ANA) using indirect immunofluorescence assay (IIFA), linear immunoassay (LIA), chemiluminescence microparticle immunoassay (CMIA), multiple microbead immunoassay (MBI) and to compare these four methods in the performance of diagnosing systemic lupus erythematosus (SLE). Serum ANA were detected in 147 SLE cases and 42 healthy controls (HCs). The sensitivity, specificity, accuracy, positive predictive value and agreement, the area under the curve of four methods in diagnosing were calculated. Finally, a diagnostic model through logistic regression was constructed. The sensitivity of CMIA and IIFA in diagnosing SLE was 89.08% and 89.12%, higher than other two methods (P < .01), while highest specificity lied in CMIA (95.24%) and LIA (95.24%). The accuracy was highest in CMIA (91.01%), and lowest in LIA (83.07%). CMIA and the other three methods had good agreement, especially with LIA (κ = .798, 95% CI, 0.708-0.88). ANA-IIFA (OR = 1.016, P < .001) and anti-SSA antibodies (OR = 1.017, P = .043) were finally included in the SLE diagnostic model, with AUC value of 0.964 (95% CI, 0.936-0.991). SLE patients exhibited 14 various ANA patterns, especially AC-1, AC-4, and AC-5. Antibodies against SSA and dsDNA were mostly seen with AC-1 and AC-4 patterns, while antibodies against RNP, Sm, SSA, dsDNA, nucleosome and PO were most frequently observed with AC-5 pattern in SLE. CMIA method is a reliable screening test for detections of antibodies related to SLE. Using ANA-IIFA and anti-SSA antibodies by CMIA can discriminate SLE patients from HCs effectively.     

A solid-phase chemiluminescence immunoassay for determination of IgG in eluates of psoriatic scales

A solid-phase chemiluminescence immunoassay for determining immunoglobulin G (IgG) in eluates of psoriatic scales is described. Polystyrene balls coated with protein A bound the IgG present in the eluates and in human sera. The bound IgG was then detected by a luminol-coupled goat anti-human IgG conjugate, characterized for antibody, protein and luminol content. This method correlated well with radial immunodiffusion in determining IgG levels in human sera (r = 0.97). The sensitivity of the assay was 0.5 μg IgG. Eluates prepared from psoriatic scales treated sequentially with buffers of decreasing pH all contained IgG when tested by this method. pH 2 eluates, however, contained more IgG than pH 3 eluates, suggesting the presence of specifically bound antibodies.     

Graves病和桥本病甲状腺细胞凋亡与Fas表达的研究

为了研究正常、Graves病 (GD )和桥本甲状腺炎 (HT )甲状腺细胞凋亡和凋亡相关蛋白Fas的变化特征及其临床意义 ,采用细胞培养方法和流式细胞术检测正常、GD和HT甲状腺细胞凋亡率和Fas表达量。结果发现 :(1)GD和HT甲状腺细胞凋亡率明显高于正常甲状腺细胞 (P <0 0 1)。其中 ,尤以HT甲状腺细胞凋亡增加最为显著 ;(2 )HT甲状腺细胞Fas表达阳性率明显高于正常和GD甲状腺细胞 (P <0 0 1) ,而GD与正常对照相比无统计学差异。以上结果表明 ,自身免疫性甲状腺疾病 (AITD )患者甲状腺细胞存在细胞凋亡和Fas表达的异常变化 ,尤以HT为显著 ,提示Fas介导的细胞凋亡参与AITD的发病过程 ,可能与HT甲状腺细胞破坏有关。     

化学发光法检测肝纤维化血清学指标的临床应用

目的探讨化学发光法检测肝纤维化指标包括三型前胶原N端肽（PⅢNP）、Ⅳ型胶原（CⅣ）、层粘蛋白（LN）及透明质酸（HA）对肝纤维化的诊断价值。方法用化学发光法检测132例乙型病毒性肝炎患者血清中肝纤维化四项指标的水平,评价四项指标鉴别诊断肝炎及肝硬化的价值。结果乙型肝炎组、肝癌组、肝硬化组血清中的四项指标依次升高,均与正常对照组差异有统计学意义（P〈0.01）。ROC曲线显示,PⅢNP、CIV、LN、HA的曲线下面积分别为0.680、0.825、0.716、0.825,灵敏度分别为39.1%、87.0%、87.0%、82.6%,特异性分别为89.8%、69.3%、53.4%、86.4%,准确性分别为79.09%、72.73%、78.18%、85.45%。在联合检测中,HA＋PⅢNP组合的灵敏度为83.0%,特异性为80.0%,准确性为80.1%。结论单个指标检测,HA是四项指标中鉴别诊断肝炎与肝硬化最有价值的一项,通过HA联合其他指标进行分析,其中HA＋PⅢNP组合特异性、准确性最高。