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。

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。

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。

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。

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。

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。

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。

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。

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。

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。

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。

BACKGROUND & AIMS: :An enzyme-linked immunosorbent assay (ELISA) was developed which permits the assay of specific secretory immunoglobulin A (sIgA) antibodies against hepatitis B surface antigen (HBsAg) in saliva. The assay is based on the binding of sIgA antibodies present in saliva to microtitre plates coated with excess of F(ab')2 anti-secretory component antibodies, followed by the addition of specific antigen, HBsAg and finally peroxidase-labelled anti-HBsAg. The assay is fast, simple, reproducible and antigen specific as shown by total absence of inhibition of specific antigen by unrelated antigens but significant inhibition of labelled anti-HBsAg by unlabelled anti-HBsAg. The values obtained for hospital personnel exposed to hepatitis infections (0.068 +/- 0.083 U/ml) and for post-icteric hepatitis B patients (0.062 +/- 0.033 U/ml) were significantly higher than values in control subjects (0.013 +/- 0.006 U/ml).

背景与目标： ：开发了一种酶联免疫吸附测定（ELISA），可以测定唾液中针对乙型肝炎表面抗原（HBsAg）的特异性分泌型免疫球蛋白A（sIgA）抗体。该测定基于唾液中存在的sIgA抗体与包被有过量F（ab'）2抗分泌成分抗体的微量滴定板的结合，然后添加特异性抗原HBsAg，最后添加过氧化物酶标记的抗HBsAg。该测定是快速，简单，可重现的和抗原特异性的，如完全不存在不相关抗原对特异性抗原的抑制，但未标记抗-HBsAg对标记抗-HBsAg的显着抑制所示。暴露于肝炎感染的医院工作人员获得的值（0.068 /-0.083 U / ml）和黄疸型乙型肝炎患者获得的值（0.062 /-0.033 U / ml）显着高于对照受试者的值（0.013 /-0.006 U / ml）。

BACKGROUND & AIMS: :The anti-interferon-gamma (IFN-gamma) autoantibody is a known cause of opportunistic non-tuberculous mycobacterial (NTM) infection in adults. Diagnosis of those patients is difficult due to the low sensitivity of bacterial culture, and because detection of the neutralizing autoantibody needs special laboratory devices. We conducted a retrospective review of indirect and inhibitory ELISA, both used for detection of anti-IFN-gamma auto-antibody in 102 patients with lymphadenopathies. We assessed hospital records of NTM isolation and/or diagnosis of NTM infection. The review revealed the compatible sensitivity and superior specificity and predictive values for inhibitory ELISA over against indirect ELISA-the latter achieving 100% specificity and positive predictive value for diagnosis of NTM infection in patients with lymphadenopathies. The results confirm functional assays that show plasma samples from NTM-infected patients with positive results by either indirect and/or inhibitory ELISA are IFN-gamma neutralizing autoantibodies. The inhibitory titer of anti-IFN-gamma auto-antibody can be used to distinguish patients with active from inactive NTM infection. Inhibitory ELISA is thus a practical, rapid, high performance tool for routine detection of anti-IFN-gamma autoantibody and NTM infection diagnosis before confirmation, enabling a timely therapeutic strategy for active infection treatment.

背景与目标： ：抗干扰素-γ（IFN-γ）自身抗体是成人机会性非结核分枝杆菌（NTM）感染的已知原因。由于细菌培养的敏感性低，并且由于中和自身抗体的检测需要特殊的实验室设备，因此难以诊断这些患者。我们对间接和抑制性ELISA进行了回顾性回顾，两者均用于检测102例淋巴腺病患者的抗IFN-γ自身抗体。我们评估了NTM隔离和/或NTM感染诊断的医院记录。该综述揭示了抑制性ELISA与间接ELISA兼容的敏感性，优越的特异性和预测值-间接ELISA对于淋巴腺病患者的NTM感染诊断具有100％的特异性和阳性预测值。该结果证实了功能测定，该测定表明通过间接和/或抑制性ELISA从NTM感染患者获得的血浆样品为IFN-γ中和自体抗体。抗IFN-γ自身抗体的抑制效价可用于区分活动性和非活动性NTM感染患者。因此，抑制性ELISA是一种实用，快速，高性能的工具，可在确认之前进行常规检测抗IFN-γ自身抗体和NTM感染的诊断，从而为及时进行主动感染治疗提供了可行的治疗策略。

BACKGROUND & AIMS: :Tau protein is a major component of paired helical filaments (PHFs) which constitute the characteristic neurofibrillary tangle lesions observed in Alzheimer's disease. Two tau mAbs have been produced which show distinct patterns of immunoreactivity with intact human tau and with tau incorporated in PHFs. The mAb 423 recognises PHFs but not human tau on immunoblots whereas mAb 7/51 reacts with human tau but its epitope is buried within the PHF and is only exposed after formic acid treatment. A competitive ELISA has been developed for both of these mAbs and these have been used to quantify the two distinct tau epitopes in PHFs. Samples containing antigen are incubated with horseradish peroxidase-conjugated mAb at 4 degrees C for 16 h and non-adsorbed antibody then measured by binding, at 37 degrees C for 1 h, to a fragment of tau coated on microtitre plates. Bound enzyme-labelled antibody is measured kinetically using a spectrophotometer capable of automatically mixing the samples throughout a 2-min incubation with substrate and chromogen. The interfacing of the plate reader with a computer permits competitive curves to be plotted automatically using Softmax. Curves are fitted using a 4-parameter logistic algorithm which allows one to determine the relative immunoreactivity for different samples. The application of these assays to monitoring biochemical fractions and quantifying distinct immunochemical presentations of tau protein with these two mAbs is described.

背景与目标： ：Tau蛋白是成对的螺旋丝（PHF）的主要成分，这些螺旋丝构成了在阿尔茨海默氏病中观察到的特征性神经原纤维缠结损伤。已生产出两种tau单克隆抗体，它们与完整的人类tau和掺入PHF的tau表现出不同的免疫反应模式。 mAb 423在免疫印迹上识别PHF，但不能识别人tau，而mAb 7/51与人tau反应，但其表位被掩埋在PHF中并且仅在甲酸处理后才暴露。已经针对这两种单克隆抗体开发了竞争性ELISA，并将它们用于定量PHF中两个不同的tau表位。将含有抗原的样品与辣根过氧化物酶偶联的单克隆抗体在4°C下孵育16小时，然后通过在37°C下与包被在微量滴定板上的tau片段结合来测量未吸附的抗体。结合酶标记的抗体是使用分光光度计动力学测量的，该分光光度计能够在与底物和色原体孵育2分钟的过程中自动将样品混合。酶标仪与计算机的连接允许使用Softmax自动绘制竞争曲线。使用4参数逻辑算法拟合曲线，该算法可以确定不同样品的相对免疫反应性。描述了这些测定法在监测生化成分和定量用这两种单克隆抗体对tau蛋白的不同免疫化学表现中的应用。

BACKGROUND & AIMS: :To improve compliance with a gluten-free diet in coeliac disease a simple prototype test kit was developed to detect gluten in foods for use at home. The test is based on monoclonal antibodies to heat-stable gluten proteins which crossreact appropriately with barley and rye proteins. It is suitable for use with a wide range of raw or cooked foods. The food is extracted with dilute hydrochloric acid and 1 drop of the extract transferred to an antibody-coated tube; enzyme-labelled gluten detection antibody is added and after 3 min the tube is washed and colour developer is added. The reaction is stopped after 2 min, stabilising the blue colour. The home kit was compared with a quantitative laboratory kit, and the qualitative agreement was very good. The kit could distinguish foods with trace gluten contents (acceptable for a "gluten-free" diet) from those with a slightly higher but unacceptable gluten content. In a trial of the prototype kit by 47 coeliac disease patients of diverse ages and educational backgrounds, 93% of tests correctly identified foods as acceptable or unacceptable.

背景与目标： ：为了改善乳糜泻中无麸质饮食的依从性，开发了一种简单的原型测试套件来检测家用食品中的麸质。该测试基于抗热稳定的面筋蛋白的单克隆抗体，该蛋白可与大麦和黑麦蛋白发生适当的交叉反应。它适用于各种生或熟食品。用稀盐酸提取食物，并将1滴提取物转移到涂有抗体的试管中。加入酶标记的麸质检测抗体，并在3分钟后清洗试管并加入显色剂。 2分钟后停止反应，稳定蓝色。将家用套件与定量实验室套件进行了比较，并且定性协议非常好。该试剂盒可以将麸质含量较低的食品（“无麸质”饮食可接受）与麸质含量稍高但不可接受的食品区分开。在由47位年龄和教育背景不同的乳糜泻患者进行的原型工具包试验中，有93％的测试正确地将食物标识为可接受或不可接受。