• 【通过点ELISA检测HIV阳性/ AIDS患者和自愿献血者口服液中的抗HIV-1 / 2抗体。】 复制标题 收藏 收藏
    DOI: 复制DOI
    作者列表:Abrão Ferreira PR,Gabriel R,Furlan TM,de Camargo Soares A,Myazaky ME,Bordim JO,Castelo A,Lewi DS
    BACKGROUND & AIMS: :Serology is the primary means for identifying patients with HIV infection and Acquired Immunodeficiency Syndrome (AIDS). Testing of serum by serologic methods has been extensively used since 1985, not only for clinical diagnosis but also for epidemiological surveillance and donor screening in blood banks. Fast serological diagnostic techniques are now being developed, using urine and oral fluid, as an alternative for anti-HIV antibody screening, and many parallel studies are proving its accuracy. The purpose of this study was to evaluate the sensitivity, specificity, accuracy, positive predictive value (PPV) and negative predictive value (NPV) of the ImmunoComb II HIV 1&2 Saliva((R)) test from Orgenics (Dot-ELISA) compared to the routine exams (ELISA and Western Blot) of HIV positive/AIDS patients, undergoing antiretroviral treatment or not, in different stages of the disease's evolution, and compared to serologic testing of known HIV negative patients by the use of serum ELISA (blood donors). To accomplish this, patients of the Immunogenic Deficiencies Control Center (CCDI) and voluntary blood donors of the Blood Bank Center of the Medical School of Sã:o Paulo/Federal University of São Paulo (EPM-UN I FESP) were evaluated. Sensitivity of Dot-ELISA in oral fluid was 100%, specificity 97.08%, PPV 96.66% and NPV 100%. The method used in this case study was shown to be highly sensitive and specific, being useful particularly in epidemiological surveillance and screening.
    背景与目标: :血清学是鉴定HIV感染和后天免疫机能丧失综合症(AIDS)患者的主要手段。自1985年以来,通过血清学方法进行血清检测已广泛用于临床诊断,还用于血库中的流行病学监测和供体筛查。现在正在开发一种快速的血清学诊断技术,使用尿液和口腔液体作为抗HIV抗体筛查的替代方法,许多平行研究证明了其准确性。这项研究的目的是评估Orgenics(Dot-ELISA)进行的ImmunoComb II HIV 1&2 Saliva(R)测试的敏感性,特异性,准确性,阳性预测值(PPV)和阴性预测值(NPV),与在疾病发展的不同阶段中接受或未接受抗逆转录病毒治疗的HIV阳性/ AIDS患者的常规检查(ELISA和Western Blot),并与使用血清ELISA(献血者)的已知HIV阴性患者的血清学检测进行比较。为此,对S&atilde:o Paulo /圣保罗联邦大学医学院(EPM-UN I FESP)的免疫原性缺乏症控制中心(CCDI)的患者和血液库中心的自愿献血者进行了评估。 Dot-ELISA在口腔液中的敏感性为100%,特异性为97.08%,PPV为96.66%,NPV为100%。该案例研究中使用的方法显示出高度的敏感性和特异性,特别适用于流行病学监测和筛查。
  • 【抗人IFN-α8特异性单克隆抗体的建立及其在酶联免疫吸附测定(ELISA)中的应用。】 复制标题 收藏 收藏
    DOI:10.1089/jir.2007.0121 复制DOI
    作者列表:Ushio C,Ariyasu H,Kayano T,Ohta H,Aga M,Ariyasu T,Ohta T,Kurimoto M,Fukuda S
    BACKGROUND & AIMS: :In the present study, we describe the generation of a series of anti-interferon-alpha8 (IFN-alpha8)-specific monoclonal antibodies (mAbs), their characterization, and the establishment of a sandwich enzyme-linked immunosorbent assay (ELISA) system for human IFN-alpha8. The sandwich ELISA system is highly sensitive to human natural IFN-alpha8 (nIFN-alpha8), with a minimum detection limit of 50 pg/mL, which did not cross-react with the other IFN preparations and several cytokines tested. Using this ELISA system, pharmacokinetic properties of an IFN-alpha preparation administered in mice were examined. We found that IFN-alpha8 has higher vascular permeability and stability than IFN-alpha2 in the circulation. These results suggest that this ELISA would be very useful for determination of IFN-alpha8 protein concentrations in various experimental samples and also of pharmacokinetic properties of IFN-alpha preparations in human.
    背景与目标: :在本研究中,我们描述了一系列抗干扰素-α8(IFN-α8)特异性单克隆抗体(mAbs)的产生,其表征以及夹心酶联免疫吸附测定(ELISA)系统的建立对于人IFN-α8。夹心ELISA系统对人天然IFN-α8(nIFN-alpha8)高度敏感,最低检测限为50 pg / mL,与其他IFN制剂和几种测试的细胞因子没有交叉反应。使用该ELISA系统,检查了在小鼠中施用的IFN-α制剂的药代动力学特性。我们发现,IFN-alpha8在循环中比IFN-alpha2具有更高的血管通透性和稳定性。这些结果表明,该ELISA对于测定各种实验样品中的IFN-α8蛋白浓度以及人中的IFN-α制剂的药代动力学特性将非常有用。
  • 3 IGRA-ELISA for Tuberculosis Diagnosis. 复制标题 收藏 收藏

    【用于肺结核诊断的IGRA-ELISA。】 复制标题 收藏 收藏
    DOI:10.7754/Clin.Lab.2019.190830 复制DOI
    作者列表:Mungmunpuntipatip R,Wiwnaitkit V
    BACKGROUND & AIMS: -2
    背景与目标: -2
  • 【鉴定羊卵附肢相关蛋白(AAAP)以开发间接ELISA及其应用。】 复制标题 收藏 收藏
    DOI:10.1186/s13071-017-2297-z 复制DOI
    作者列表:Wang Z,Yang J,Niu Q,Brayton KA,Luo J,Liu G,Yin H,Liu Z
    BACKGROUND & AIMS: BACKGROUND:Ovine anaplasmosis is a tick-borne disease that is caused by Anaplasma ovis in sheep and goats. The pathogen is widely distributed in tropical and subtropical regions of the world. At present, diagnosis of the disease mainly depends on microscopy or nucleic acid based molecular tests, although a few serological tests have been applied for the detection of A. ovis infection. RESULTS:Here we describe the identification of an A. ovis protein that is homologous to the A. marginale appendage-associated protein (AAAP). We expressed a recombinant fragment of this protein for the development of an indirect enzyme-linked immunosorbent assay (ELISA) for the detection of A. ovis. Anaplasma ovis-positive serum showed specific reactivity to recombinantly expressed AAAP (rAAAP), which was further confirmed by the rAAAP ELISA, which also demonstrated no cross-reactivity with sera from animals infected with A. bovis or other related pathogens in sheep and goats. Testing antibody kinetics of five experimentally infected sheep for 1 year demonstrated that the rAAAP ELISA is suitable for the detection of early and persistent infection of A. ovis infections. Investigation of 3138 field-collected serum samples from 54 regions in 23 provinces in China demonstrated that the seroprevalence varied from 9.4% to 65.3%, which is in agreement with previous reports of A. ovis infection. CONCLUSIONS:An A. ovis derived antigenic protein, AAAP, was identified and the antigenicity of the recombinant AAAP was confirmed. Using rAAAP an indirect ELISA assay was established, and the assay has been proven to be an alternative serological diagnostic tool for investigating the prevalence of ovine anaplasmosis of sheep and goats.
    背景与目标: 背景:绵羊羊膜无病是一种由传播的疾病,是由绵羊和山羊的无形体引起的。该病原体广泛分布在世界热带和亚热带地区。目前,该疾病的诊断主要取决于显微镜或基于核酸的分子检测,尽管一些血清学检测已被用于检测猪链球菌感染。
    结果:在这里,我们描述了一种与牛角耳附属物相关蛋白(AAAP)同源的牛卵杆菌蛋白的鉴定。我们表达了此蛋白的重组片段,用于开发间接酶联免疫吸附测定(ELISA)来检测羊肠曲霉。羊膜羊水阳性血清对重组表达的AAAP(rAAAP)有特异性反应,rAAAP ELISA进一步证实了该反应,它也证实与感染牛羊链球菌或绵羊和山羊的其他相关病原体的血清无交叉反应。测试五只经实验感染的绵羊的抗体动力学,历时1年,证明rAAAP ELISA适用于检测早期和持续感染的A. ovis感染。对中国23个省54个地区的3138份野外采集的血清样本进行的调查表明,血清阳性率从9.4%到65.3%不等,这与以前的A. ovis感染报告相符。
    结论:鉴定出了一种源自农杆菌的抗原蛋白AAAP,并证实了重组AAAP的抗原性。使用rAAAP建立了间接ELISA测定法,该测定法已被证明是用于调查绵羊和山羊绵羊羊浆虫病患病率的另一种血清学诊断工具。
  • 【使用单克隆抗体的ELISA检测肺炎支原体抗原。】 复制标题 收藏 收藏
    DOI: 复制DOI
    作者列表:Hirschberg L,Holme T
    BACKGROUND & AIMS: :Seven monoclonal antibodies directed against major antigens of Mycoplasma pneumoniae were selected for the development of an antigen detection assay. Three of these were directed to the 170,000-dalton adhesin of M. pneumoniae. The test was an antigen-capture enzyme immunoassay using the different monoclonal antibodies for capture of antigen and a polyclonal rabbit antiserum as detection reagent. With three of the monoclonal antibodies a detection limit of approximately 2 ng M. pneumoniae protein was obtained, as determined by titration of M. pneumoniae organisms in buffer. The detection limit of the assays was only slightly less when the other four monoclonal antibodies were used. In artificially infected nasopharyngeal aspirates the detection limit was approximately 10 times lower. The fact that no significant differences in the detection limit of the assays were recorded using monoclonal antibodies directed against different antigens indicates that these antigens were available for reaction with antibodies irrespective of their location in intact M. pneumoniae cells. In the assay there were no significant cross-reactions with a number of bacterial species potentially colonizing the respiratory tract, except for a protein A-positive strain of Staphylococcus aureus. Our test is equally sensitive to another recently described ELISA using polyclonal antibodies. In comparison with other recommended methods such as immunoblot and culture-amplified antigen detection assays, the ELISA is more rapid and less laborious.
    背景与目标: :选择了七种针对肺炎支原体主要抗原的单克隆抗体,用于开展抗原检测测定。其中三个直接针对肺炎支原体的170,000道尔顿黏附素。该测试是一种抗原捕获酶免疫分析,使用不同的单克隆抗体捕获抗原并使用多克隆兔抗血清作为检测试剂。通过在缓冲液中滴定肺炎支原体,可以使用三种单克隆抗体获得约2 ng的肺炎支原体蛋白检测限。当使用其他四种单克隆抗体时,测定的检测限仅略低。在人工感染的鼻咽抽吸物中,检出限降低了约10倍。使用针对不同抗原的单克隆抗体未检测到检测限的显着差异这一事实表明,这些抗原可用于与抗体反应,而不论它们在完整的肺炎支原体细胞中的位置如何。在该测定中,除金黄色葡萄球菌的蛋白A阳性菌株外,没有与可能在呼吸道中定殖的多种细菌发生显着的交叉反应。我们的测试对最近使用多克隆抗体的另一种ELISA同样敏感。与其他推荐的方法(例如免疫印迹和培养物扩增的抗原检测测定法)相比,ELISA更快,更省力。
  • 【RIDASCREEN®汉坦病毒Puumala IgG / IgM ELISA分析的分析性能。】 复制标题 收藏 收藏
    DOI:10.3390/v12020226 复制DOI
    作者列表:Depypere M,Lagrou K,Van Esbroeck M,Houben E,Van Ranst M
    BACKGROUND & AIMS: :The National Reference Center for Hantavirus in Belgium is currently using the Hantavirus IgM/IgG ELISA Progen kit (Heidelberg, Germany) for the detection of the most prevalent Hantavirus in Western Europe, Puumala virus (PUUV). Two commercially available PUUV kits were compared: Progen and RIDASCREEN® Hantavirus Puumala IgM/IgG ELISA assay (Darmstadt, Germany). METHODS:The sensitivity was evaluated with a panel of 68 samples from patients with an acute infection (n = 44) or a past infection (n = 24). Specificity was evaluated with a panel of 62 samples from patients with potentially false borderline results (n = 7) (no seroconversion), seronegative samples (n = 25) and potentially cross reacting samples (n = 30). Discordances were resolved by immunoblot. Substantial agreement was calculated using Cohen kappa coefficient. RESULTS:The RIDASCREEN® kit showed a higher specificity (IgM: 94.3%; IgG: 94.4%) than the Progen kit (IgM: 77.0% IgG: 93.0%). The sensitivity for IgM ELISA was 100% for both assays. IgG sensitivity was, respectively, 98.3% and 100% for Progen and RIDASCREEN®. A Cohen kappa coefficient of 0.76 and 0.90 was found between Puumala IgM and IgG, respectively. CONCLUSIONS:This study showed a higher specificity for the RIDASCREEN® kit than the Progen kit, while the sensitivity was as good as for the Progen kit.
    背景与目标: :比利时国家汉坦病毒参考中心目前正在使用汉坦病毒IgM / IgG ELISA Progen试剂盒(德国海德堡)来检测西欧最流行的汉坦病毒,即Puumala病毒(PUUV)。比较了两种市售的PUUV试剂盒:Progen和汉达病毒Puumala IgM / IgG ELISA分析(德国达姆施塔特)。
    方法:对一组来自急性感染(n = 44)或既往感染(n = 24)患者的68份样本进行了敏感性评估。评估了一组62个样本的特异性,这些样本来自可能有错误的临界结果(n = 7)(无血清转化),血清阴性样本(n = 25)和潜在的交叉反应样本(n = 30)的患者。不一致之处通过免疫印迹解决。使用Cohen kappa系数计算出基本一致性。
    结果:试剂盒显示出比Progen试剂盒(IgM:77.0%IgG:93.0%)更高的特异性(IgM:94.3%; IgG:94.4%)。两种测定的IgM ELISA灵敏度均为100%。 Progen和RIDASCREEN®的IgG敏感性分别为98.3%和100%。 Puumala IgM和IgG之间的Cohen kappa系数分别为0.76和0.90。
    结论:这项研究表明,RIDASCREEN®试剂盒的特异性高于Progen试剂盒,而灵敏度与Progen试剂盒一样好。
  • 【空气过敏原分析及其临床意义。 I.通过RAST抑制,Mab-ELISA,嗜碱性粒细胞组胺释放和逆流免疫电泳对过敏原进行免疫化学定量。】 复制标题 收藏 收藏
    DOI:10.1111/j.1398-9995.1991.tb00611.x 复制DOI
    作者列表:Johnsen CR,Abrahamsen L,Stahl Skov P,Johansen N,Poulsen LK
    BACKGROUND & AIMS: :The aim was to compare IgE and IgG4 RAST-inhibition assay (RI), monoclonal antibody ELISA (Mab-ELISA), counter current immuno electrophoresis (CCIE) and histamine release from basophil leukocytes (HR) for allergen quantification with special reference to aeroallergen detection. As components of indoor aeroallergens, cat, dog, and Derm. pter. allergen extracts were selected for the experiments. To evaluate unspecific interference, these allergens were compared mutually and with Cladosporium herbarum. Allergen extracts in varying dilutions were mixed with crushed glass fibre filter materials, eluted, recovered by centrifugation, and allergen concentration quantified by the assays. Equal sensitivity was found for both IgE- and IgG4-RI assaying cat allergen (in the range 5-50 SQ-U/ml) and dog allergen (in the range 10(2)-10(3) SQ-U/ml). The IgG4-RI assaying Derm. pter. was more sensitive (50 SQ-U/ml) than IgE-RI (2*10(3) SQ-U/ml). The ranges of allergen detection limits for the Mab-ELISA were equal for cat and Derm. pter. (10-10(2) SQ-U/ml). The range of allergen detection limits for CCIE, assaying dog were 10(4)-10(5) SQ-U/ml. The ranges of allergen detection limits for HR were equal for cat and Derm. pter. (10-10(2) SQ-U/ml), and 10(2)-10(3) SQ-U/ML for dog. Because of cross-reactivity, a minor degree of interference was observed in the IgE-RI and the HR test for the highest concentration of cat and dog allergens.
    背景与目标: :目的是比较IgE和IgG4 RAST抑制测定(RI),单克隆抗体ELISA(Mab-ELISA),逆流免疫电泳(CCIE)和嗜碱性白细胞(HR)释放的组胺以定量过敏原,并特别参考空气过敏原检测。作为室内空气过敏原的成分,猫,狗和皮肤。 pter。选择过敏原提取物用于实验。为了评估非特异性干扰,将这些变应原相互比较并与桔梗比较。将不同稀释度的过敏原提取物与碎玻璃纤维过滤材料混合,洗脱,离心分离回收,并通过化验定量量化过敏原浓度。发现IgE-和IgG4-RI检测猫过敏原(范围为5-50 SQ-U / ml)和狗过敏原(范围为10(2)-10(3)SQ-U / ml)均具有相同的敏感性。 IgG4-RI测定皮肤。 pter。比IgE-RI(2 * 10(3)SQ-U / ml)更敏感(50 SQ-U / ml)。 Mab-ELISA的过敏原检出限的范围对于猫和真皮均相等。 pter。 (10-10(2)SQ-U / ml)。 CCIE的过敏原检出限范围为10(4)-10(5)SQ-U / ml。猫和真皮的HR过敏原检出限范围是相等的。 pter。 (10-10(2)SQ-U / ml)和10(2)-10(3)SQ-U / ML用于狗。由于交叉反应,在IgE-RI和HR测试中观察到猫和狗过敏原的最高浓度的干扰程度较小。
  • 【基于外壳蛋白基因的柑桔类三链病毒不对称PCR-ELISA分型方法的建立。】 复制标题 收藏 收藏
    DOI:10.1016/j.jviromet.2008.09.030 复制DOI
    作者列表:Nolasco G,Santos C,Silva G,Fonseca F
    BACKGROUND & AIMS: :The coat protein gene of isolates of citrus tristeza virus (CTV) from 20 citrus-producing regions around the world was amplified by RT-PCR, TA cloned, and characterized by SSCP. Haplotypes that produced different patterns within each geographic region were sequenced and a database of 153 accessions of CTV was assembled. Phylogenetic analysis revealed the existence of seven well-defined clusters (Coefficient of differentiation 0.78). An asymmetric PCR-ELISA typing (APET) assay was developed in the frame of this clustering pattern using a set of eight hybridisation probes. The membership of any unknown haplotype is determined by comparing its pattern of reaction against the whole set of probes and not, as previously done in hybridisation assays, in an all-or-nothing basis. Interpretation of the results is objective and done through a visual basic application that compares the rates of hydrolysis of the ELISA substrate of an assayed isolate to a matrix of rates of hydrolysis obtained from standard haplotypes. This assay was validated and showed a better ability to resolve haplotypes than other assays to which it was compared experimentally. It may be automated to the same extent as any ELISA.
    背景与目标: :通过RT-PCR扩增了来自全球20个柑橘产区的柑橘变形杆菌病毒(CTV)分离株的外壳蛋白基因,TA克隆并通过SSCP进行了鉴定。对在每个地理区域内产生不同模式的单倍体进行测序,并收集了153份CTV的数据库。系统发育分析显示存在七个明确定义的簇(分化系数0.78)。使用一组八种杂交探针,在这种聚类模式的框架内开发了一种不对称PCR-ELISA分型(APET)测定法。通过将其反应模式与整套探针进行比较来确定任何未知单倍型的成员,而不是像以前在杂交测定中所做的那样,不是全有还是全无。结果的解释是客观的,并且是通过视觉基础应用程序完成的,该应用程序将测定的分离物的ELISA底物的水解速率与从标准单倍型获得的水解速率矩阵进行比较。该试验已经过验证,与其他试验进行了比较,它显示出更好的分辨单倍型的能力。它可以自动化到与任何ELISA相同的程度。
  • 9 ELISA system for human endothelial lipase. 复制标题 收藏 收藏

    【人内皮脂肪酶的ELISA系统。】 复制标题 收藏 收藏
    DOI:10.1373/clinchem.2012.187914 复制DOI
    作者列表:Ishida T,Miyashita K,Shimizu M,Kinoshita N,Mori K,Sun L,Yasuda T,Imamura S,Nakajima K,Stanhope KL,Havel PJ,Hirata K
    BACKGROUND & AIMS: BACKGROUND:Endothelial lipase (EL) regulates the metabolism of HDL cholesterol (HDL-C). However, the role of EL in regulating plasma HDL-C concentrations and EL's potential involvement in atherosclerosis in humans has not been fully investigated due to the lack of reliable assays for EL mass. We developed an ELISA system for serum EL mass. METHODS:Human recombinant EL proteins, purified from cultured media of human EL-transfected Chinese hamster ovary cells, were used as antigen and calibrator. Two specific monoclonal antibodies were generated in mice against recombinant EL protein for a sandwich ELISA. We measured EL mass in human serum using EL recombinant protein as a calibration standard. RESULTS:The EL antibodies did not cross-react with lipoprotein lipase and hepatic triglyceride lipase. The detection limit of the ELISA was 20 pg/mL, which is approximately 10 times lower than that of previous ELISA systems. Recovery of spiked EL in serum was 90%-105%. Assay linearity was intact with a >4-fold dilution of serum. Intra- and interassay CVs were <5%. The serum EL mass in 645 human subjects was [mean (SE)] 344.4 (7.7) pg/mL (range 55.2-1387.7 pg/mL). Interestingly, serum EL mass was increased in patients with diagnosed cardiovascular disease and inversely correlated with serum HDL-C concentrations. There was no difference in EL mass between pre- and postheparin plasma samples. CONCLUSIONS:This ELISA should be useful for clarifying the impact of EL on HDL metabolism and EL's potential role in atherosclerosis.
    背景与目标: 背景:内皮脂肪酶(EL)调节HDL胆固醇(HDL-C)的代谢。但是,由于缺乏可靠的EL质量检测方法,尚未充分研究EL在调节血浆HDL-C浓度中的作用以及EL在人类动脉粥样硬化中的潜在作用。我们开发了用于血清EL质量的ELISA系统。
    方法:从人EL转染的中国仓鼠卵巢细胞培养基中纯化得到的人重组EL蛋白作为抗原和校准物。在小鼠中产生了针对重组EL蛋白的两种特异性单克隆抗体,用于三明治ELISA。我们使用EL重组蛋白作为校准标准,测量了人血清中的EL含量。
    结果:EL抗体与脂蛋白脂肪酶和肝甘油三脂脂肪酶无交叉反应。 ELISA的检出限为20 pg / mL,比以前的ELISA系统低约10倍。血清中加标EL的回收率为90%-105%。血清稀释度> 4倍时,测定线性保持不变。批内和批间CVs小于5%。 645位人类受试者的血清EL质量为[平均值(SE)] 344.4(7.7)pg / mL(范围55.2-1387.7 pg / mL)。有趣的是,确诊为心血管疾病的患者血清EL量增加,并且与血清HDL-C浓度呈负相关。肝素治疗前后血浆样品的EL质量无差异。
    结论:该ELISA方法可用于阐明EL对HDL代谢的影响以及EL在动脉粥样硬化中的潜在作用。
  • 【通过各种曲霉菌生产黄曲霉毒素和环吡嗪酸:ELISA分析。】 复制标题 收藏 收藏
    DOI:10.1007/BF03192259 复制DOI
    作者列表:Huang X,Dorner JW,Chu FS
    BACKGROUND & AIMS: :An enzyme-linked Immunosorbent assay (ELISA) was used to monitor a total of 153 fungi in theAspergillus flavus group, Including 130A. flavus, 15A. parasiticus and 8A. tamarii, for their ability to produce aflatoxins (AFs) and cyclopiazonic acid (CPA) in a mycologlcal broth-sucrose-yeast extract medium. Of 15A. parasiticus isolates, ten produced AFs In a range of 12.4 to 89.3 μg/vial (average 56.9 μg/vial); two isolates produced only trace amounts of AFs and three isolates produced none at all. Production of CPA was not demonstrated in anyA. parasiticus isolate. On the other hand, all A. tamarii isolates produced only CPA with a range of 310 to 1100 gmg/vial. Fifteen percent (14.6%) of theA. flavus isolates (19/130) produced more than 500 μg CPA/vial, but yielded no or little AF (less than 0.1 μg/vial). About 22.3% ofA. flavus (29/130) that produced less than 500 μg of CPA also yielded little or no aflatoxin. MostA. flavus isolates (44.6%) produced both CPA (50 to 300 μg/vial) and AFs (10 to 40 μg/vial). About 9.2% of theA. flavus are low CPA producers (less than 100 μg/vial) but yielded higher amounts of AFs. A small percentage (12/130 or 9.2%) of A. flavus isolates produced neither CPA nor aflatoxin. Excluding the isolates that produced neither AFs nor CPA, there is a negative correlation between the production of CPA and AFs by most A.flavus isolates. Data obtained from ELISA for the production of CPA were consistent with TLC results. Thus, the ELISA method for CPA and AFB could be applied to the screening of toxigenic fungi. Data on the simultaneous production of both toxins by a large percentage of the toxigenicA. flavus isolates suggest that there is a potential health hazard for co-existence of both toxins in foods and feeds.
    背景与目标: :酶联免疫吸附试验(ELISA)用于监测黄曲霉组中总共153种真菌,包括130A。黄褐色,15A。寄生虫和8A。 tamarii,因为它们在霉菌肉汤-蔗糖-酵母提取物培养基中产生黄曲霉毒素(AFs)和环吡嗪酸(CPA)的能力。 15A。副寄生物分离物,产生十个AF,范围为12.4至89.3μg/小瓶(平均56.9μg/小瓶);两个分离株仅产生痕量的AF,而三个分离株则完全不产生。在任何A中都没有证明CPA的产生。寄生虫分离物。另一方面,所有的塔氏曲霉分离株仅产生CPA,范围为310至1100 gmg /小瓶。占A的百分之十五(14.6%)。黄酮分离物(19/130)产生的CPA /小瓶超过500μg,但没有产生或产生很少的AF(小于0.1μg/小瓶)。约占A的22.3%。产生少于500μgCPA的黄酮(29/130)也产生很少或没有黄曲霉毒素。多数。黄酮分离物(44.6%)产生CPA(50至300μg/小瓶)和AF(10至40μg/小瓶)。约占A的9.2%。黄褐斑是低CPA生产者(少于100μg/瓶),但产生的AF数量更高。一小部分(12/130或9.2%)黄曲霉分离株既不产生CPA也不产生黄曲霉毒素。除去既不产生AF也不产生CPA的分离株,大多数黄曲霉分离株在CPA和AF的产生之间存在负相关。从ELISA中获得的用于生成CPA的数据与TLC结果一致。因此,可将CPA和AFB的ELISA方法应用于产毒真菌的筛选。关于大量毒素同时产生两种毒素的数据。黄酮分离物表明,食物和饲料中两种毒素的共存均存在潜在的健康危害。
  • 【评估捕获ELISA以检测针对Wegener肉芽肿病中针对蛋白酶3的抗中性粒细胞胞质抗体:一项多中心研究的第一个结果。】 复制标题 收藏 收藏
    DOI:10.1093/rheumatology/keh028 复制DOI
    作者列表:Csernok E,Holle J,Hellmich B,Willem J,Tervaert C,Kallenberg CG,Limburg PC,Niles J,Pan G,Specks U,Westman K,Wieslander J,De Groot K,Gross WL
    BACKGROUND & AIMS: OBJECTIVE:To evaluate the performance characteristics of direct and capture ELISA for the detection of PR3-ANCA in Wegener's granulomatosis (WG) in international ANCA reference laboratories. METHODS:Serum samples were derived from patients with histological and clinical diagnosis of WG (n = 60), rheumatoid arthritis (RA) (n = 30) and healthy controls (n = 30). Each of them was tested for the presence of ANCA by indirect immunofluorescence technique (IFT), direct and capture ELISA in six international reference laboratories (Massachusetts General Hospital, Boston; Wieslab AB, Lund; University of Maastricht; University Hospital Groningen; Mayo Clinic, Rochester; Rheumaklinik Bad Bramstedt/University of Schleswig-Holstein Campus Lübeck). Each centre tested the sera according to their house protocols of IFT and ELISA. The diagnostic performance of each test was estimated by receiver operating characteristic curve analysis and sensitivity and specificity in detection of ANCA/PR3-ANCA were calculated for the respective methods. RESULTS:In patients histologically and clinically known as WG, the detection of ANCA by IFT varied between 52 and 83% among the participating centres. PR3-ANCA positivity with the different ELISAs ranged from 53 to 80% in direct ELISA and from 72 to 76% in capture ELISA. While most capture ELISAs successfully detected PR3-ANCA, there were significant differences between IFT and direct ELISA results between laboratories. ROC curve analysis demonstrated that in five of six laboratories the overall diagnostic performance of capture ELISA was superior to IFT and direct ELISA, respectively. CONCLUSION:Capture ELISA is a highly sensitive assay for detection of PR3-ANCA in WG and should be used in conjunction with compatible clinical picture and histological evidence.
    背景与目标: 目的:评估直接和捕获ELISA在国际ANCA参考实验室中检测韦格纳肉芽肿病(WG)中PR3-ANCA的性能特征。
    方法:血清样本来自具有组织学和临床诊断的WG(n = 60),类风湿关节炎(RA)(n = 30)和健康对照组(n = 30)的患者。通过间接免疫荧光技术(IFT),直接和捕获ELISA在六个国际参考实验室(波士顿马萨诸塞州总医院,隆德Wieslab AB,马斯特里赫特大学,格罗宁根大学,格罗宁根大学医院,梅奥诊所,罗切斯特; Rheumaklinik Bad Bramstedt /石勒苏益格-荷尔斯泰因大学吕贝克校区)。每个中心都根据自己的IFT和ELISA协议对血清进行测试。通过接收器工作特性曲线分析评估每个测试的诊断性能,并针对相应方法计算出ANCA / PR3-ANCA检测的灵敏度和特异性。
    结果:在组织学和临床上被称为WG的患者中,IFT对ANCA的检出率在参与研究的中心之间在52%至83%之间。在直接ELISA中,不同ELISA的PR3-ANCA阳性率在53%至80%之间,在捕获ELISA中,PR3-ANCA阳性率在72%至76%之间。尽管大多数捕获ELISA成功检测到PR3-ANCA,但实验室间IFT和直接ELISA结果之间存在显着差异。 ROC曲线分析表明,在六个实验室中的五个实验室中,捕获ELISA的总体诊断性能分别优于IFT和直接ELISA。
    结论:捕获ELISA是一种用于检测WG中PR3-ANCA的高灵敏度检测方法,应与兼容的临床图片和组织学证据结合使用。
  • 【用于测量牛软骨离体模型中活性ADAMTS-4的竞争性ELISA的开发和表征。】 复制标题 收藏 收藏
    DOI:10.1016/j.matbio.2012.12.001 复制DOI
    作者列表:He Y,Zheng Q,Simonsen O,Petersen KK,Christiansen TG,Karsdal MA,Bay-Jensen AC
    BACKGROUND & AIMS: :ADAMTS-4 (aggrecanase1) is believed to play an important role in the degradation of aggrecan during the progression of joint diseases. ADAMTS-4 is synthesized as a latent pro-enzyme that requires the removal of the pro-domain, exposing the N-terminal neoepitope, to achieve activity. We developed a monoclonal antibody against this neoepitope of active ADAMTS-4. Furthermore, we established and characterized a competitive ELISA for measuring active ADAMTS-4 form applying the specific antibody. We used this assay to profile the presence of active ADAMTS-4 and its aggrecan degradation product (NITEGE(373)) in a bovine cartilage ex vivo model. We found that after stimulation with catabolic factors, the cartilage initially released high levels of aggrecanase-derived aggrecan fragments into supernatant but subsequently decreased to background levels. The level of active ADAMTS-4 released into the supernatant and retained in the cartilage matrix increased continuously throughout the 21days of the study. The activity of ADAMTS-4 on the last day of catabolic stimulation was verified in vitro by adding deglycosylated or native aggrecan to the conditioned medium. Samples of human cartilage affected by varying degrees of osteoarthritis stained strongly for active ADAMTS-4 where surface fibrillation and clustered chondrocytes were observed. This assay could be an effective tool for studying ADAMTS-4 activity and for screening drugs regulating ADAMTS-4 activation. Moreover, it could be a potential biomarker for degenerative joint disease.
    背景与目标: :ADAMTS-4(aggrecanase1)被认为在关节疾病进展期间在聚集蛋白聚糖的降解中起重要作用。 ADAMTS-4被合成为潜在的原酶,需要去除原结构域,暴露N末端新表位才能达到活性。我们开发了针对活性ADAMTS-4的这种新表位的单克隆抗体。此外,我们建立并鉴定了一种竞争性ELISA,用于测量应用特异性抗体的活性ADAMTS-4形式。我们使用此测定法分析了牛软骨离体模型中活性ADAMTS-4及其蛋白聚糖降解产物(NITEGE(373))的存在。我们发现,在分解代谢因子刺激后,软骨最初将高水平的软骨聚集蛋白聚糖酶衍生的软骨聚集蛋白片段释放到上清液中,但随后降至背景水平。在整个研究的21天中,释放到上清液中并保留在软骨基质中的活性ADAMTS-4的水平持续增加。通过在条件培养基中添加去糖基化的或天然的聚集蛋白聚糖,体外验证了分解代谢刺激的最后一天ADAMTS-4的活性。受不同程度的骨关节炎影响的人类软骨样品对活跃的ADAMTS-4进行了强烈染色,其中观察到了表面纤颤和聚集的软骨细胞。该测定法可能是研究ADAMTS-4活性和筛选调节ADAMTS-4活化的药物的有效工具。此外,它可能是退行性关节疾病的潜在生物标志物。
  • 【人胰腺脂肪酶相关蛋白2:沿消化道的组织定位,并使用特异性ELISA在胰腺液中定量。】 复制标题 收藏 收藏
    DOI:10.1016/j.bbagen.2006.06.005 复制DOI
    作者列表:Eydoux C,Aloulou A,De Caro J,Grandval P,Laugier R,Carrière F,De Caro A
    BACKGROUND & AIMS: :Human pancreatic lipase-related protein 2 (HPLRP2) was previously found to be secreted by the exocrine pancreas. HPLRP2 shows a high level of activity on galactolipids, and might be involved in the digestion of these common vegetable lipids. Specific antibodies were raised in rabbits using a synthetic HPLRP2 peptide selected for its weak amino acid homology with the corresponding peptides of classical human pancreatic lipase (HPL) and human pancreatic lipase-related protein 1 (HPLRP1). ELISA and Western blotting data showed that these antibodies did not react with HPL or HPLRP1. Various tissues from the digestive tract were subjected to Western blotting analysis with the specific anti-peptide HPLRP2 antibody and the expression of HPLRP2 was detected in the pancreas and colon. An ELISA was developed for specifically measuring the HPLRP2 levels in pure pancreatic juice. This procedure was performed using the anti-peptide HPLRP2 antibody as the captor antibody and a biotinylated anti-HPLRP2 polyclonal antibody as the detector antibody. The lowest HPLRP2 quantification limit was found to be 50 microg/L and the reference range for the present assay was 50 microg-500 microg/L. HPL and HPLRP2 levels were measured using specific ELISAs in pancreatic juice from patients with and without pancreatic disorders. Patients with chronic calcifying pancreatitis (CCP) had significantly lower levels of both HPL and HPLRP2 than the controls subjects. The mean HPLRP2 to HPL ratio was estimated to be 28.30% (w/w) and 23.96% (w/w) in controls subjects and CCP patients, respectively, and the difference was not significant. The levels of HPL and HPLRP2 are therefore similarly reduced in both healthy patients and CCP patients.
    背景与目标: :人胰脂肪酶相关蛋白2(HPLRP2)先前被发现是由外分泌胰腺分泌的。 HPLRP2对半乳糖脂具有很高的活性,可能与这些常见植物脂质的消化有关。使用合成的HPLRP2肽在兔中产生特异性抗体,该肽的选择是由于其与经典人胰脂肪酶(HPL)和人胰脂肪酶相关蛋白1(HPLRP1)的相应肽具有弱氨基酸同源性。 ELISA和蛋白质印迹数据表明,这些抗体不与HPL或HPLRP1反应。用特异性抗肽HPLRP2抗体对消化道的各种组织进行Western印迹分析,并在胰腺和结肠中检测到HPLRP2的表达。已开发出一种ELISA用于专门测量纯胰液中HPLRP2的水平。使用抗肽HPLRP2抗体作为捕获抗体,并使用生物素化的抗HPLRP2多克隆抗体作为检测器抗体进行此过程。发现最低HPLRP2定量限为50微克/升,本测定的参考范围为50微克至500微克/升。使用特异的ELISA法测量患有和不患有胰腺疾病的患者的胰液中的HPL和HPLRP2水平。慢性钙化性胰腺炎(CCP)患者的HPL和HPLRP2水平均明显低于对照组。对照组和CCP患者的平均HPLRP2与HPL比率分别估计为28.30%(w / w)和23.96%(w / w),差异不显着。因此,在健康患者和CCP患者中,HPL和HPLRP2的含量均降低。
  • 【用于急诊科临床肺栓塞的ELISA D-二聚体测量:为期一年的安全性和医生处方观察性研究。】 复制标题 收藏 收藏
    DOI:10.1179/acb.2003.58.4.004 复制DOI
    作者列表:Fr V,Hainaut P,Fr T,Elamly A,Dessomme B,Lavenne E,Reynaert MS
    BACKGROUND & AIMS: OBJECTIVES:To validate the safety profile of a rapid ELISA D-dimer as the first diagnostic step in the clinical suspicion of pulmonary embolism (PE) in outpatients admitted to an emergency department (ED), and to retrospectively evaluate the appropriateness of the physician's prescription. DESIGN AND SETTING:An observational study of all patients admitted to the ED of an urban university teaching hospital with signs and symptoms justifying the prescription of a rapid ELISA D-dimer measurement (Vidas; Biomerieux; France) as the first line diagnostic test for PE. Acute PE was established or excluded according to an appropriate combination of the D-dimer concentration, the lung scintigraphy, the spiral computerized tomography (spiral CT), the venous ultrasonography, and the arteriography in case of uncertain results. All patients with D-dimer values under the cut-off point of 500 ng/ml were followed up after 6 months. RESULTS:395 patients were studied. A normal D-dimer concentration < 500 ng/ml was found in 179 patients (45% of the cohort). The retrospective analysis showed that none of these patients were found to have a high pre-test clinical probability. None of these 179 patients received anticoagulation nor displayed a PE event during a 6-month period (negative predictive value 100%; 95% CI, 98.0 to 100%; sensitivity 100%; 95% CI, 90.3 to 100%). Among the 216 patients (55%) with D-dimer values above 500 ng/ml, PE was confirmed in 32 cases, for a prevalence of the disease of 8.1%. Eighty-six patients (22%) had no additional testing in spite of positive D dimer values > 500 ng/ml, pointing out a 22% rate of inappropriate use of the D-dimer measurement. CONCLUSION:This observational study confirms that a normal rapid ELISA D-dimer value (< 500 ng/ml) used as a first diagnostic step in ruling out the diagnosis of PE is a safe clinical practice when the pre-test clinical probability is low or intermediate. Nevertheless, the low prevalence rate of the disease (8.1%) suggests a potential overused and inappropriate prescription.
    背景与目标: 目的:验证快速ELISA D-二聚体的安全性,将其作为急诊科(ED)门诊患者在临床怀疑肺栓塞(PE)的第一步诊断步骤,并回顾性评估医师处方的适用性。
    设计与地点:对某城市大学教学医院急诊科住院患者的体征和症状进行观察性研究,该体征和症状证明了快速ELISA D-二聚体测定处方的正确性(维达斯; Biomerieux;法国),是体育一线诊断测试。根据结果​​的不确定性,根据D-二聚体浓度,肺闪烁显像,螺旋计算机断层扫描(螺旋CT),静脉超声和动脉造影的适当组合,建立或排除急性PE。所有D-二聚体值低于500 ng / ml临界点的患者均在6个月后进行了随访。
    结果:对395例患者进行了研究。在179名患者中发现了正常的D-二聚体浓度<500 ng / ml(占队列的45%)。回顾性分析表明,这些患者中没有人具有很高的预检临床可能性。这179名患者在6个月内均未接受抗凝治疗或未发生PE事件(阴性预测值100%; 95%CI为98.0至100%;敏感性为100%; 95%CI为90.3至100%)。 D-二聚体值高于500 ng / ml的216例患者(55%)中,有32例确诊为PE,该病患病率为8.1%。尽管D二聚体值大于500 ng / ml,但仍有86例患者(22%)没有进行其他检查,指出22%的D-二聚体测量值使用不当。
    结论:这项观察性研究证实,当检测前临床概率低或检测到PE时,正常的快速ELISA D-二聚体值(<500 ng / ml)作为排除PE诊断的第一步诊断步骤是安全的临床实践。中间的。然而,该疾病的低患病率(8.1%)表明存在潜在的过度使用和不适当的处方。
  • 【使用不同的净化步骤通过ELISA和TLC方法测定的保加利亚玉米样品中伏马菌素B1的出现率。】 复制标题 收藏 收藏
    DOI:10.1007/BF02946696 复制DOI
    作者列表:Vrabcheva T,Stroka J,Anklam E
    BACKGROUND & AIMS: :An enzyme linked immunosorbent assay (ELISA) method and a thin-layer chromatography (TLC) method with two different clean-up procedures have been used to analyse maize samples (food and feeding stuff) obtained from Bulgaria. The occurrence of Fumonisin B1 in Bulgarian maize was found to be in the range of what was reported for other European regions with a mean value of 1.5 mg/kg, 2.1 mg/kg and 1.8 mg/kg when analysed by ELISA and thin-layer chromatography with solid phase extraction clean up and immunoaffinity clean up, respectively. It was shown that results of the different methods are comparable, indicating that the methods applied are sufficiently reliable tools for the simple and rapid screening of maize samples.
    背景与目标: :酶联免疫吸附测定(ELISA)方法和薄层色谱(TLC)方法具有两种不同的净化程序,已用于分析从保加利亚获得的玉米样品(食品和饲料)。通过酶联免疫吸附测定(ELISA)和薄层分析法发现保加利亚玉米中伏马菌素B1的发生在其他欧洲地区的报道范围内,平均值为1.5 mg / kg,2.1 mg / kg和1.8 mg / kg固相萃取层析和免疫亲和层析。结果表明,不同方法的结果具有可比性,表明所采用的方法是用于简单,快速筛查玉米样品的足够可靠的工具。

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