间接免疫荧光法阴性的抗核抗体标本中特异性抗体检测的临床意义

抗核抗体(ANA)在临床中应用于自身免疫性疾病(AID)的诊断已有50多年的历史。抗核抗体是以细胞成分为靶抗原的器官非特异性自身抗体的总称,以Hep-2细胞为抗原基质的间接免疫荧光法(IIF)被认为是ANA检测的"金标准"[1]。除间接免疫荧光法以外,近年来随着技术的不断发     

蛋白芯片技术及免疫印迹技术检测抗核抗体的比对研究

目的通过比对免疫印迹法与蛋白芯片法对血清标本中抗核抗体的检测结果,验证这两种方法是否具有等效性。方法分别用免疫印迹技术和蛋白芯片技术检测70例临床标本的抗核抗体,记录检验结果,并对检查结果进行统计学分析。结果经配对资料卡方检验分析,两种方法测定SSA、SSB、SM、Scl-70、Jo-1和CENP时结果差异无统计学意义(P>0.05)。仅在测定RNP时结果差异有统计学意义(P=0.04)。结论两种检测方法所测抗核抗体的结果一致性较好,蛋白芯片技术可用于临床抗核抗体的检测。     

抗ENA抗体与抗核抗体的相关性

目的探讨抗核抗体（ANA）荧光核型与抗可提取性核抗原抗体（ENA）之间的相关性。方法ANA采用间接免疫荧光法检测,抗ENA抗体采用欧蒙免疫斑点法检测,检测结果采用双盲对照法分析。结果312例抗ENA抗体阳性病例ANA的阳性率为93．3％,荧光核型主要以颗粒型为主,抗Scl-70抗体阳性时则以核仁型为主,抗Jo-1抗体阳性的ANA荧光核型比较分散。另外检测的124例ANA阳性病例中,抗ENA抗体常规6项阳性率为71．8％。结论抗ENA抗体类型与ANA荧光核型之间没有绝对的规律,临床检测自身抗体时需将两者结合分析。     

间接免疫荧光法筛查抗核抗体与特异性抗体检测的相互关系

目的:对间接免疫荧光法(IIF)筛查抗核抗体与特异性抗体检测之间的相互关系进行研究分析。对其临床意义予以确定。方法:抽取1894份临床连续送检血清标本的抗核抗体,将这1894例标本随机分成三组,分别作为自身免疫性疾病组、疑似自身免疫性疾病组和非自身免疫性疾病组,采用间接免疫荧光法对这些标本进行筛查,采用线性免疫印迹法(LIA)对抗核抗体进行检测,对检测结果的相互关系进行分析与总结。结果:在所调查的结果中,两种方法均阳性的有833份,约占43.98%;IIF检查结果表现出阳性而LIA检查结果为阴性的有246份,约占12.99%;IIF检查结果表现出阴性而LIA检查结果为阳性的有189份,约占9.98%;两种方法均呈阴性的有626份,约占33.05%。两种方法的总体符合率为77.03%,以上同级的数据均具有明显的统计学差异(P<0.05)。结论:采用间接免疫荧光法筛查抗核抗体与特异性抗体检测的结果显示出,两种方法具有很高的符合率,这对临床来说具有重要意义。     

酶联免疫双抗原夹心法检测梅毒螺旋体抗体

在我国近年来梅毒的发病率有逐年上升的趋势 ,研制出快速、简便、灵敏且特异的诊断方法是当前的主要研究方向。梅毒螺旋体感染机体后可产生两种抗体 ,一种是抗心磷脂抗体 ,即非特异性抗体 ,亦称反应素。主要检测方法有性病研究实验室试验 (VDRL )和血浆反应素环状卡片试验 (RPR     

肝抗原自身抗体检测在自身免疫性肝病诊断中的意义

目的:探讨自身免疫性肝脏疾病的自身抗体特征,以提高对该病的认识。方法:在85例患者中,自身免疫性肝炎(AIH)17例,原发性胆汁性肝硬化(PBC)22例,原发性硬化性胆管炎(PSC)5例,AIH/PBC重叠综合征11例,不明原因肝损伤30例。采用间接免疫荧光法和免疫印迹法分别检测抗核抗体(ANA)、抗线粒体抗体(AMA)、抗平滑肌抗体(SMA)、抗线粒体Ⅱ型抗体(AMA-M2)、抗肝肾微粒体抗体Ⅰ型(LKM-1)、抗肝胞浆I型抗体(LC-1)和抗可溶性肝抗原/肝胰抗原(SLA/LP)等。结果:自身免疫性肝炎组ANA、SMA、抗LKM-1、抗LC-1、抗SLA/LP的阳性率分别为82.3%、52.9%、11.8%、5.9%、11.8%。原发性胆汁性肝硬化组ANA、AMA、AMA-M2阳性率为86.4%、100%、95.5%。原发性硬化性胆管炎组检测出ANA2例。AIH/PBC重叠组检测出ANA7例,AMA6例,SMA2例。不明原因肝损伤组检测出ANA9例,AMA1例。结论:多数自身免疫性肝病伴有特异性自身抗体的出现,肝抗原自身抗体的检测将有助于自身免疫性肝病患者的诊断及治疗。     

HIV1/2抗体与P24抗原联合检测的意义研究

目的 研究HIV1/2抗体与P24抗原联合检测对临床诊断的影响.方法 自2006年1月-2011年3月分别收集ELISA法检测HIV1/2抗体吸光度(OD值)为临界值及大于等于阳性对照的患者血清33、29份,采用电化学发光法联合检测HIV1/2抗体与P24抗原(HIVCOM).追踪患者免疫印迹HIV确证试验结果,研究HIV1/2抗体与P24抗原联合检测对临床诊断的影响.结果 33例临界值吸光度患者中27例HIV1/2抗体与P24抗原阳性,6例阴性,29例大于等于阳性对照值吸光度的患者中28例阳性,1例阴性,免疫印迹HIV确证试验的检测结果与联合检测HIV1/2抗体及P24抗原结果一致.结论 HIV1/2抗体与P24抗原联合检测较ELISA法能够更早更准确的发现HIV感染,对临床的与早期诊断和治疗具有重要的意义及应用价值.     

用鼠类腹水癌细胞检测抗核抗体的临床意义

抗核抗体 (ANA)是以细胞核为靶细胞的自身抗体的总称 ,抗核抗体的检测对某些疾病尤其是自身免疫疾病诊断是肯定的。对疾病愈后判定十分有价值。常用检测方法为免疫荧光法和免疫酶法 ,长期以来检测中所用抗原片为鼠肝片或用人来源(HeP -2 )培养细胞[1,2 ],因来源困难在基层难以开展。我们试用大鼠艾氏 (Ehrlich)腹水癌细胞为ANA的抗原片 ,经与灵敏度高的鼠肝片、人来源HeP -2细胞比较 ,发现灵敏度高、结果清晰。下面我们对 58例临床确认患者以大鼠腹水癌细胞ANA抗原片与灵敏度高的鼠肝片、HeP -2细胞做检测及比较 ,现将试验结果报告如…     

登革热抗原抗体联合检测与双抗体检测效能比较

目的 比较抗原抗体联合检测与双抗体快速检测2种方法检测登革热的效能,为登革热快速诊断提供依据。方法 收集登革热临床诊断病例血清样本449份(病例组)、流行病学和(或)临床上判断与登革热无关血清样本689份(阴性对照组),分别进行抗原抗体联合检测(IgM、IgG和NS1抗原)及双抗体检测(IgG和IgM),以临床诊断为金标准,对2种方法检测结果进行描述分析和一致性分析。结果 纳入研究的449例病例组和689例阴性对照组中,联合检测阳性率(34.1%)高于双抗检测(29.7%),且均低于临床诊断阳性率(39.4%),差异均有统计学意义(χ²值分别为20.61、7.03和51.33,P值均<0.01)。联合检测结果与实际临床诊断一致性(kappa=0.863)高于双抗检测(kappa=0.745),联合与双抗检测法结果一致性较好(kappa=0.729)。阴性对照组有9份样本经联合和(或)双抗检测法检测为登革热阳性,其中4例(3例发热待排查病例,1例体检人员)2种方法均为阳性,增补为登革热临床诊断病例。结论 抗原抗体联合胶体金检测登革热不仅方便、快捷、经济,与临床...     

幽门螺杆菌抗体分型检测在临床中的应用分析

目的：在胃炎、消化道溃疡患者中进行幽门螺杆菌(Hp)抗体分型检测,分析Hp毒力分型结果,辅助临床医生更好地开展“选择性根除”Hp。方法：应用免疫印迹法对510例胃炎、消化道溃疡患者检测血清Hp 4种抗体(CacA抗体、VacA抗体、UreA抗体、UreB抗体),根据抗体结果进行Hp分型并进行统计分析。结果：350例(阳性率68.63%)患者检出Hp抗体,男性阳性率vs女性阳性率无统计学意义(P>0.05)。Ⅰ型Hp抗体(CagA和/或VacA抗体阳性)阳性率52.75%显著高于Ⅱ型Hp抗体(CagA和Vac A抗体阴性)阳性率15.88%(χ²=153.7,P<0.001),CagA抗体阳性率(50.98%)高于VacA抗体阳性率(38.82%)(χ²=77.493, P<0.001),UreB抗体阳性率(68.24%)高于UreA(40.78%)(χ²=15.233, P<0.001),差异均有统计学意义。结论：应用免疫印迹法检测Hp 4种抗体,能够为Hp感染进行分型,为临床制定Hp诊疗方案提供...     

水体中苯并芘检测试剂盒研制及效果评价

目的 研制间接竞争酶联免疫吸附法(ELISA)检测苯并芘试剂盒。方法 采用棋盘滴定法确定抗原抗体的最佳用量;采用间接ELISA分析法确定苯并芘检测的最适条件,包括pH值、盐离子浓度及甲醇浓度。结果 抗原最佳包被浓度为10 μg/mL,单克隆抗体稀释倍数为 1:5000;在最优pH值为7.4、盐离子浓度为0.01 mol/L及甲醇浓度为10%的条件下,检测方法在5~50 ng/mL范围内线性相关,线性方程为y=-28.105 Ln(x)+123.66,R²=0.9937,IC₅₀为13.74 ng/mL,苯并芘的最低检出限为3.3 ng/mL;在水样中的加标回收率为94.8%~112.3%;变异系数为4.38%~9.72%。结论 该方法适用于水体中苯并芘的检测,可用于大批量样品的快速筛查。     

应用膜抗原荧光抗体试验检测人群水痘-带状疱疹病毒血清抗体阳性率

目的 应用膜抗原荧光抗体试验(FAMA)方法调查广州地区正常人群水痘-带状疱疹病毒(VZV)的流行现状.方法 采用以VZV感染细胞作为抗原、异硫氰酸荧光素(FITC)标记的羊抗人IgG作为二抗的FAMA试验,对随机抽取的592份正常人血清标本进行特异性VZV抗体检测.结果 FAMA试验检测VZV抗体与其他的疱疹类病毒的相应抗体不产生交叉反应.应用此法检测592份血清标本中,VZV抗体总体阳性率为76.52%;1～、4～、7～、14～、20～、30～、40～及≥50岁年龄组血清抗体阳性率分别是14.67%、51.56%、73.91%、91.26%、92.78%、95.65%、98.11%和100%.1～3岁年龄组血清抗体阳性率最低,血清抗体阳性率随年龄的增大而升高;不同性别间阳性率差异无统计学意义(P0.005),不同年龄组间阳性率差异有统计学意义(P<0.001).结论 采用FAMA法检测人群中VZV抗体与其他疱疹类病毒的相应抗体不产生交叉反应,是VZV抗体检测的可靠方法 .应以1～3岁儿童为VZV疫苗免疫的首选对象.     

Establishing China's national standards of antigen content and neutralizing antibody responses for evaluation of enterovirus 71 (EV71) vaccines

Enterovirus 71 (EV71) is a highly infectious agent that causes hand-foot-mouth disease (HFMD) in humans. Effective vaccination against EV71 infection is critically important, given the recent outbreak of HFMD in the Asia-Pacific region, where it has shown significant mortality and morbidity. There is currently no approved anti-viral therapy available to treat the disease. While several vaccine manufacturers are actively developing EV71 vaccines, there are no international reference standards available to conduct quality control on EV71 vaccines or to assess the effectiveness of EV71 vaccines in immunized populations. In the current report, antigen reference standard based on the C4 subtype of the EV71 vaccine strain was developed. In addition, neutralizing antibody (NTAb) reference panels were analyzed and standards with various neutralizing titers were selected. These reference antigens were used to calibrate vaccine samples from several producers and found that five EV71 antigens and the national reference standards showed good linearity and parallelism. Moreover, mice immunized with various vaccines at doses standardized by these national references showed comparable NTAb responses. Finally, the national NTAb reference panels were found to effectively reduce assay discrepancy between different labs. Taken together, these national reference standards are highly valuable for the standardization and evaluation of EV71 vaccines.     

Adjuvant effect of biodegradable poly(DL-lactic acid) granules capable for antigen release following intraperitoneal injection

Ovalbumin (OVA)-containing poly(DL-lactic acid) (PDLLA) granules were prepared with different conditions. Following the intraperitoneal (i.p.) immunization of mice with the granules containing OVA, production of anti-OVA IgG antibody in the mouse serum was investigated. The i.p. injection of the granules induced a strong antibody production compared with that of free OVA, irrespective of the amount of OVA released for initial a few weeks and the period of OVA release. The serum level of IgG antibody induced by the granules was retained at a high level over 16 weeks although the period of OVA release and the amount of OVA released initially were different from each other. The initial OVA release for a few weeks was essential to induce the enhanced antibody production. Comparison of mice immunization by granules with different OVA loadings but at a similar dose revealed that antibody level was higher for the granules with lower loading than for those with the higher loading. However, when the granules were injected after encapsulation into a poly(vinyl alcohol) (PVA) hydrogel tube, the difference in their antibody level became insignificant. Because PVA encapsulation did not affect the OVA release profile, this finding indicates that the injection amount of the granules seems to have influenced the antibody production. We conclude that the release profile of OVA is not always a key factor to enhance the antibody production of OVA-containing granules so far as the initial OVA controlled release is achieved.     

Evaluation of the effect of maternally derived antibody on response to MMR vaccine in Thai infants

BackgroundAlthough the number of measles cases declined globally in response to anti-measles immunisation campaigns, measles has re-emerged. A review of current vaccination policies is required to improve measles elimination strategies.MethodsA pseudotype-based virus neutralisation assay (PVNA) was used to measure neutralising antibody titres in serum samples collected from Thai infants at six timepoints before and after two-doses of MMR (1&2) vaccination (ClinicalTrials.gov no. NCT02408926). Vesicular stomatitis virus (VSV) luciferase pseudotypes bearing the haemaglutinin (H) and fusion (F) glycoproteins of measles virus (MeV) were prepared. Serial dilutions of serum samples were incubated with VSV (MeV) pseudotypes and plated onto HEK293-human SLAM1 cells; the neutralising antibody titre was defined as the dilution resulting in 90% reduction in luciferase activity.ResultsNeutralising antibody titres in infants born with high levels of maternal immunity (H group) persisted at the time of the first MMR vaccination, and those infants did not respond effectively by developing protective titres. In contrast, infants with lower maternal immunity (L group) developed protective titres of antibody following vaccination. Responses to the second MMR vaccination were significantly higher (P = 0.0171, Wilcoxon signed-rank test) in the H group. The observed correlation between anti-MeV IgG level and neutralising antibody titre in Thai infants indicates the possibility of using rapid IgG testing as a surrogate measure for neutralising activity to define clinical protection levels within populations.ConclusionThese results demonstrate that varying the timing of the first MMR immunisation according to the level of acquired maternal immunity could increase vaccination immunogenicity and hence accelerate measles eradication.     

Cost-effectiveness of monoclonal antibody and maternal immunization against respiratory syncytial virus (RSV) in infants: Evaluation for six European countries

BackgroundRespiratory syncytial virus (RSV) imposes a substantial burden on pediatric hospital capacity in Europe. Promising prophylactic interventions against RSV including monoclonal antibodies (mAb) and maternal immunizations (MI) are close to licensure. Therefore, we aimed to evaluate the cost-effectiveness of potential mAb and MI interventions against RSV in infants, for six European countries.MethodsWe used a static cohort model to compare costs and health effects of four intervention programs to no program and to each other: year-round MI, year-round mAb, seasonal mAb (October to April), and seasonal mAb plus a catch-up program in October. Input parameters were obtained from national registries and literature. Influential input parameters were identified with the expected value of partial perfect information and extensive scenario analyses (including the impact of interventions on wheezing and asthma).ResultsFrom the health care payer perspective, and at a price of €50 per dose (mAb and MI), seasonal mAb plus catch-up was cost-saving in Scotland, and cost-effective for willingness-to-pay (WTP) values ≥€20,000 (England, Finland) or €30,000 (Denmark) per quality adjusted life-year (QALY) gained for all scenarios considered, except when using ICD-10 based hospitalization data. For the Netherlands, seasonal mAb was preferred (WTP value: €30,000-€90,000) for most scenarios. For Veneto region (Italy), either seasonal mAb with or without catch-up or MI was preferred, depending on the scenario and WTP value. From a full societal perspective (including leisure time lost), the seasonal mAb plus catch-up program was cost-saving for all countries except the Netherlands.ConclusionThe choice between a MI or mAb program depends on the level and duration of protection, price, availability, and feasibility of such programs, which should be based on the latest available evidence. Future research should focus on measuring accurately age-specific RSV-attributable hospitalizations in very young children.     

Identification of the critical attribute(s) of EBV gp350 antigen required for elicitation of a neutralizing antibody response in vivo

Vaccine prophylaxis with EBV glycoprotein 350 (gp350) subunit plus adjuvant has been demonstrated clinically to protect individuals against infectious mononucleosis (IM), but the specifications of the antigen required to elicit this protection has remained largely theoretical. Previous studies have shown that antibodies to gp350 comprise the principle component of EBV-neutralizing sera. Further, a murine monoclonal antibody against gp350 (clone 72A1) is able to prevent infection by the virus both in vitro and in vivo. In the present study, we identify the 72A1 epitope on recombinant gp350 antigen as the site required for binding to CD21 on human B cells. We also identify the need for conformational-dependence of the antigen to generate EBV-neutralizing antibodies in vivo. Further, we have characterized the glycosylation status and antigenicity profiles of both native and denatured CHO-produced soluble gp350 as well as non-glycosylated protein produced in Escherichia coli. Collectively our in vitro and in vivo data demonstrate the requirement for a conformationally accessible 72A1 epitope on gp350 to elicit EBV-neutralizing responses, and establish this as a critical attribute of this vaccine antigen. These data provide direction for commercial vaccine development, as the absence of this epitope on either E. coli-expressed or denatured gp350, may limit production and purification options for the antigen.     

妊娠期补钙预防妊娠高血压综合征的Meta分析

研究补钙对妊娠高血压综合征的预防作用。应用 Meta分析方法对我国 1992～ 2 0 0 2年公开发表的 13篇补钙预防妊娠高血压综合征的资料进行综合定量分析。补钙的比值比 OR为 0 .2 4 78( 0 .1792～ 0 .3373) ,补钙能使孕妇出现妊娠高血压综合征的风险平均降低 73% ,补钙有预防妊娠高血压综合征的作用     

Immunoassay procedures to detect exposure to aflatoxin B1 and benzo(a)pyrene in animals and man at the DNA level

Summary Immunological methods were used to examine human liver for the presence of aflatoxin-DNA adducts and human lung for benzo(a)pyrene diol-epoxide DNA (BPDE-DNA) adducts. Eight liver samples obtained from Czechoslovakian patients with primary hepatocellular carcinoma were studied, seven of which had detectable anti-aflatoxin inhibitory material. Values ranged between 0.63 and 3.51 picomoles aflatoxin per mg DNA. In a separate, independent study performed in another laboratory the one sample with no aflatoxin bound to DNA also had no free aflatoxin present in the liver. In the case of the human lung DNA samples, 12 samples were examined, the samples having been removed during thoracic surgery, and five had detectable anti-BPDE-DNA antibody activity. The positive samples were all from smokers and had inhibitory values ranging from 4 to 12 femtomoles per mg DNA. Samples were prepared by immunoconcentration prior to analysis. These preliminary results support the view that immunological methods can be used to examine human tissue DNA for carcinogen adducts.