BACKGROUND & AIMS: We developed a direct microtiter plate enzyme immunoassay to measure estradiol-17 beta in saliva. The assay has a commercially available monoclonal antibody, raised against estradiol-17 beta-6-carboxymethyloximebovine serum albumin, and a homologous horseradish peroxidase conjugate measured colorimetrically. The detection limit (equivalent to B0-3 SD) is 365 amol/well or 7.3 pmol/L when 50-microL samples are assayed. Cross-reactivity with estrone and estriol, testosterone, or progesterone is < 0.2%. Estradiol-17 beta was measured in daily samples over five natural menstrual cycles and eight cycles stimulated as a preliminary to in vitro fertilization, and the concentrations and fluctuations found agreed with previously published data. This method gives results in approximately 3 h and may be useful for fertility monitoring and management.

背景与目标： 我们开发了一种直接的微量滴定板酶免疫测定法，用于测量唾液中的雌二醇-17β。该测定法具有针对雌二醇-17β-6-羧甲基肟肟牛血清白蛋白的市售单克隆抗体，以及用比色法测定的同源辣根过氧化物酶结合物。当检测50微升样品时，检出限（相当于B0-3 SD）为365 amol /孔或7.3 pmol / L。与雌酮和雌三醇，睾丸激素或孕酮的交叉反应性<0.2％。在体外受精的五个自然月经周期和刺激的八个周期中，对每日样品中的雌二醇17β进行了测量，发现其浓度和波动与先前公布的数据一致。该方法可在约3小时内得出结果，可能对生育力监测和管理有用。

BACKGROUND & AIMS: :An automated microparticle enzyme immunoassay (IMx Rubella IgG Antibody Assay; Abbott Laboratories, North Chicago, Ill.) was compared with a conventional enzyme-linked immunosorbent assay (ELISA) for detection of rubella-specific immunoglobulin G (IgG) in 400 consecutive antenatal patients. There was complete agreement between the two tests in this population, which had a positivity rate of 99% for rubella-specific IgG antibodies. The performance of the IMx was also evaluated at the cutoff zone by assaying 64 selected antenatal serum samples with low or negative rubella antibody titers as determined by ELISA. Overall, the IMx was found to be a specific, sensitive assay for the detection of rubella-specific IgG and is virtually fully automated for easy performance.

背景与目标： ：将自动微粒酶免疫测定（IMx风疹IgG抗体测定；伊利诺伊州北芝加哥的雅培实验室）与常规酶联免疫吸附测定（ELISA）相结合，以检测400例连续的产前风疹特异性免疫球蛋白G（IgG）耐心。在该人群中的两次测试之间完全一致，风疹特异性IgG抗体的阳性率为99％。 IMx的性能也可以通过在64个选择的产前血清样本中通过ELISA测定的低或阴性风疹抗体滴度来评估，在临界区进行评估。总体而言，IMx被发现是一种用于检测风疹特异性IgG的特异性，灵敏的检测方法，并且实际上是全自动的，操作简便。

BACKGROUND & AIMS: :Dengue virus (DENV) is the most prevalent arbovirus in the world, found mainly in tropical regions. As clinical manifestations present frequently as nonspecific febrile illness, laboratory diagnosis is essential to confirm DENV infections and for epidemiological studies. Recombinant envelope (E) antigens of four serotypes of DENV were used to develop an immunoglobulin G enzyme-linked immunosorbent assay (IgG-ELISA). To evaluate the IgG-ELISA, a panel of serum samples that had been tested previously by a plaque reduction neutralization test (PRNT) was investigated for the presence of anti-E antibodies against the four DENV serotypes. IgG-ELISA was found to have a sensitivity (91%) and specificity (98%) at a receiver-operating characteristic (ROC) optimized cutoff and demonstrated high performance as well as good indexes. A concordance of 97% was achieved between both assays, and only 21/704 (3%) samples were not concordant. The results of the present study demonstrate a moderate correlation between neutralizing antibody titers and IgG-ELISA values. These findings indicate that the recombinant protein-based IgG-ELISA is a suitable method for routine serodiagnosis, monitoring and seroepidemiological studies of DENV infections.

背景与目标： 登革热病毒（DENV）是世界上最流行的虫媒病毒，主要发现于热带地区。由于临床表现经常以非特异性高热疾病的形式出现，因此实验室诊断对于确定DENV感染和进行流行病学研究至关重要。四种血清型DENV的重组包膜（E）抗原用于开发免疫球蛋白G酶联免疫吸附测定（IgG-ELISA）。为了评估IgG-ELISA，研究了先前通过噬斑减少中和测试（PRNT）测试过的一组血清样品中是否存在针对四种DENV血清型的抗E抗体。发现IgG-ELISA在接受者操作特征（ROC）优化的临界值上具有灵敏度（91％）和特异性（98％），并表现出较高的性能和良好的指标。两种测定之间的一致性达到97％，只有21/704（3％）个样品不一致。本研究的结果表明中和抗体滴度与IgG-ELISA值之间存在适度的相关性。这些发现表明，基于重组蛋白的IgG-ELISA是用于DENV感染的常规血清诊断，监测和血清流行病学研究的合适方法。

BACKGROUND & AIMS: BACKGROUND:Cystatin C is a low-molecular-weight protein that is freely filtered by the glomerulus and catabolized after reabsorption by the proximal tubular cells in healthy subjects. Urinary cystatin C is a potential biomarker for tubular damage including acute kidney injury (AKI) in the acute phase when patients are submitted to the intensive care unit. METHODS:The aim of this study was to perform a method validation of urinary analysis of cystatin C by particle-enhanced turbidimetric immunoassay (PETIA) on a high-throughput chemical analyzer. Total assay time was 10 min. The antigen excess, linearity, lower limit of quantification (LoQ), recovery, assay precision, stability, and interference caused by hemoglobin were evaluated. RESULTS:The LoQ was calculated to 0.020 mg/l with a coefficient of variation (CV) ≤ 10%. No hook effect was observed and the assay was linear over the studied interval less than 0.020-0.950 mg/l with a regression of R² = 0.9994. The assay had a recovery between 93-100% and the assay precision had a total CV of less than 3.5%. Cystatin C was stable for 3 days in room temperature and 14 days in +4C. The assay did not show any major interference with hemoglobin at a hemoglobin concentration of 10 g/L. The reference interval for urine cystatin C was less than 0.166 mg/l. CONCLUSION:The urinary cystatin C PETIA showed good precision and performance characteristics including short test turnaround times that are necessary qualifications for a biomarker at a routine laboratory.

背景与目标： 背景：胱抑素C是一种低分子量蛋白质，在健康受试者中被肾小球自由过滤并在近端肾小管细胞重吸收后分解代谢。当患者被送往重症监护病房时，尿半胱氨酸蛋白酶抑制剂C是潜在的肾小管损伤的生物标志物，包括急性期的急性肾损伤（AKI）。
方法：本研究的目的是通过在高通量化学分析仪上进行的颗粒增强浊度免疫测定法（PETIA）对尿液中的胱抑素C进行尿分析进行方法验证。总测定时间为10分钟。评估了抗原过量，线性，定量下限（LoQ），回收率，测定精度，稳定性和血红蛋白引起的干扰。
结果：计算出的最低定量为0.020 mg / l，变异系数（CV）≤10％。没有观察到钩效应，并且在小于0.020-0.950mg / l的研究间隔内该测定是线性的，回归值R 2 ＝ 0.9994。该测定的回收率在93-100％之间，测定精度的总CV小于3.5％。胱抑素C在室温下稳定3天，在4C下稳定14天。该测定在血红蛋白浓度为10 g / L时未显示出对血红蛋白的任何主要干扰。尿胱抑素C的参考间隔小于0.166 mg / l。
结论：尿半胱氨酸蛋白酶抑制剂PETIA显示出良好的精度和性能特征，包括较短的测试周转时间，这是常规实验室中生物标志物的必要资格。

BACKGROUND & AIMS: :We studied the use of the INNO-LIA syphilis score assay in the resolution of discordant positive screening results of the Murex ICE Syphilis enzyme immunoassay (EIA) with the confirmatory results of both the Serodia Treponema pallidum particle agglutination (TPPA) and the fluorescent treponemal antibody-absorption (FTA-Abs) assays, for the serological diagnosis of syphilis. This was an observational study on the serum samples received by the Syphilis Laboratory, Hong Kong, during the period from January 2006 to December 2012. A total of 801 serum samples with discordant positive screening EIA results were used. Consensus results of such serum samples were derived from results of the EIA, TPPA and FTA-abs assays. The age range of the individuals was 14 to 104 years (median of 52). There were 369 males and 432 females. Of 378 serum samples, 139 showed agreement among positive results, 23 of 310 showed agreement among indeterminate results and 277 of 465 showed agreement among negative results. The proportions of agreement among positive, indeterminate and negative results were 0.37 (95% CI 0.32-0.42), 0.07 (95% CI 0.05-0.11) and 0.60 (95% CI 0.55-0.64), respectively; kappa 0.55 (95% CI 0.49-0.60). There were 60 serum samples with positive consensus results but negative INNO-LIA syphilis score results and 10 with negative consensus results but positive INNO-LIA syphilis score results. Although the INNO-LIA syphilis score assay can be considered a valid alternative confirmatory test for the serological diagnosis of syphilis, the present study showed that its use in the resolution of discordant positive screening EIA results was moderate. A more extensive characterization of serum samples with discordant reactive screening treponemal test results is necessary.

背景与目标： ：我们研究了INNO-LIA梅毒评分测定在解决Murex ICE梅毒酶免疫测定（EIA）不一致的阳性筛查结果中的应用，并同时证实了梅毒螺旋体梅毒颗粒凝集（TPPA）和荧光性视网膜色素变性抗体吸收（FTA-Abs）分析，用于梅毒的血清学诊断。这是对香港梅毒实验室从2006年1月至2012年12月期间收集的血清样本的一项观察性研究。共使用了801份EIA结果阳性筛查结果不一致的血清样本。此类血清样品的共识结果来自EIA，TPPA和FTA-abs分析的结果。个人的年龄范围是14至104岁（中位数为52岁）。男369例，女432例。在378份血清样品中，有139份阳性结果一致，310份中有23份不确定结果一致，465份样品中277份阴性结果一致。阳性，不确定和阴性结果之间的一致性比例分别为0.37（95％CI 0.32-0.42），0.07（95％CI 0.05-0.11）和0.60（95％CI 0.55-0.64）; κ0.55（95％CI 0.49-0.60）。共有60份血清样本的共识结果为阳性，但INNO-LIA梅毒得分结果为阴性；有10份血清样本的共识结果为阴性，但INNO-LIA梅毒得分结果为阳性。尽管可以将INNO-LIA梅毒评分测定法视为梅毒血清学诊断的有效替代验证测试，但本研究表明，该方法在解决不一致的阳性筛查EIA结果中的应用是中等的。必须对血清样品进行更广泛的表征，并采用不协调的反应性筛检方法。

BACKGROUND & AIMS: CONTEXT:Heterophilic antibodies are a well-described interferent but poorly appreciated and are often not a recognized problem affecting most immunoassays. We describe for the first time heterophilic antibodies interference affecting an adrenocorticotropic hormone (ACTH) assay in a patient with Cushing's syndrome due to bilateral nodular adrenal hyperplasia. CASE:A 60-year-old retired female nurse underwent extensive invasive investigations, which were ultimately unnecessary, as a result of initial analytical interference in the ACTH assay, which could not be resolved using a proprietary heterophilic binding reagent. RESULTS:This case highlights the inherent difficulty of diagnosing Cushing's syndrome and the large emphasis placed on laboratory tests. The consequence of not initially identifying interference in this patient's laboratory test results led to unnecessary and costly investigations with potentially adverse outcomes. CONCLUSIONS:Clinicians and the laboratory community need to be continuously vigilant and view laboratory results with caution when they are inconsistent with the clinical picture. This approach is paramount, especially at a time of increasing automation and ever-diminishing scientist involvement in sample processing.

背景与目标： 背景：嗜异性抗体是一种被很好描述的干扰物，但人们对其了解不多，并且通常不是公认的影响大多数免疫测定的问题。我们首次描述了由于双侧结节性肾上腺皮质增生而影响库欣综合征的患者中的促肾上腺皮质激素（ACTH）测定中的嗜异性抗体干扰。
案例：一名60岁的退休女护士接受了广泛的侵入性检查，由于ACTH分析中存在最初的分析干扰，最终没有必要进行此项检查，而使用专用的异源结合试剂无法解决该问题。
结果：该病例突出了诊断库欣氏综合症的固有困难，并着重于实验室检查。最初没有识别出对该患者实验室检查结果的干扰，其结果导致不必要且昂贵的调查，并可能产生不利的后果。
结论：临床医生和实验室界需要保持警惕，并在与临床情况不一致时谨慎观察实验室结果。这种方法至关重要，尤其是在自动化程度不断提高且科学家对样品处理的参与不断减少的时候。

BACKGROUND & AIMS: BACKGROUND:The neuroprotein S100 released into the circulation has been suggested as a reliable marker for primary brain damage. However, safe identification of relevant traumatic brain injury (TBI) may possibly be hampered by S100 release from peripheral tissue. The objective of this study was to measure early S100 levels using the Elecsys S100 immunoassay for real-time assessment of severe TBI in multiple trauma. METHODS:Consecutively admitted multiple trauma patients (injury severity score >or=16 points) were stratified according to the results of the initial cerebral computed tomography (CCT) examination. S100 serum levels were determined at admission and at 6, 12, 24, 48 and 72 h after trauma. Data were correlated to creatine phosphokinase (CK) and lactate dehydrogenase (LDH) serum levels. Using receiver operating characteristic (ROC) analysis, the discriminating power of S100 measurement was calculated for the detection of CCT+ findings. RESULTS:Median S100 levels of CCT+ patients (n=9; 37 years) decreased from 3.30 microg/L at admission to 0.41 microg/L 72 h after trauma. They revealed no significant differences to CCT- patients (n=18; 44 years), but remained elevated compared to controls. Median CK and LDH levels correlated with the corresponding S100 levels during the first 24 h after trauma. ROC analysis displayed a maximum area under the curve of only 0.653 at 12 h after trauma. No significant difference was calculated for the differentiation between CCT+ and CCT- patients. CONCLUSIONS:Measurements of S100 serum levels using the Elecsys S100 immunoassay are not reliable for the real-time detection of severe TBI in multiple trauma patients. Due to soft tissue trauma or bone fractures, S100 is mainly released from peripheral sources such as adipocytes or skeletal muscle cells.

背景与目标： 背景：已被释放到循环中的神经蛋白S100被认为是原发性脑损伤的可靠标志物。但是，从周围组织释放S100可能会妨碍对相关创伤性脑损伤（TBI）的安全识别。这项研究的目的是使用Elecsys S100免疫测定法测量早期S100水平，以实时评估多发性创伤中的严重TBI。
方法：根据最初的脑计算机断层扫描（CCT）检查的结果，对连续入院的多位创伤患者（损伤严重程度评分≥16分）进行分层。在入院时以及创伤后6、12、24、48和72小时确定S100血清水平。数据与肌酸磷酸激酶（CK）和乳酸脱氢酶（LDH）血清水平相关。使用接收器工作特性（ROC）分析，计算出S100测量值的判别力以检测CCT结果。
结果：CCT患者的中位S100水平（n = 9； 37岁）从入院时的3.30微克/升降至创伤后72小时的0.41微克/升。他们显示与CCT患者无显着差异（n = 18； 44岁），但与对照组相比仍然升高。在创伤后的最初24小时内，中位CK和LDH水平与相应的S100水平相关。 ROC分析显示，创伤后12小时，曲线下的最大面积仅为0.653。对于CCT和CCT-患者之间的区别，没有计算出显着差异。
结论：使用Elecsys S100免疫测定法测量S100血清水平不能可靠地实时检测多发性创伤患者中的严重TBI。由于软组织创伤或骨折，S100主要从周围来源释放，例如脂肪细胞或骨骼肌细胞。

BACKGROUND & AIMS: :A new immunoassay technique is described which uses totally internally reflected light to excite the fluorescence of fluorescein labeled antibody which has become bound to a hapten--protein conjugate absorbed on a quartz-plate in the antibody solution. The presence of any free hapten in solution reduces the amount of antibody free to bind to the surface and thus reduces the fluorescence signal. Measurement of the decrease of the fluorescent signal then gives a measure of the concentration of free hapten present. The technique is simple, fast and has high intrinsic sensitivity and specificity. It has been demonstrated for phenyl arsonic acid and morphine. Free morphine at a concentration of 2 X 10(-7) M is readily detected.

背景与目标： ：描述了一种新的免疫测定技术，该技术利用全内反射光激发荧光素标记的抗体的荧光，该荧光素已与吸附在抗体溶液中石英板上的半抗原-蛋白质偶联物结合。溶液中任何游离半抗原的存在都会减少自由结合至表面的抗体数量，从而降低荧光信号。然后，测量荧光信号的减少，就可以测量出存在的游离半抗原的浓度。该技术简单，快速，并且具有很高的内在敏感性和特异性。已证明可用于苯a酸和吗啡。浓度为2 X 10（-7）M的游离吗啡很容易被检测到。

BACKGROUND & AIMS: A recombinant baculovirus containing a cDNA clone encoding the nucleocapsid (NP) protein of Newcastle disease virus (strain Ulster 2C) has been used to infect insect cells (Spodoptera frugiperda). High levels of overexpressed NP protein were observed, comprising up to 40% of total cellular protein, which were subsequently shown to be antigenic. Nucleoprotein derived from the crude soluble lysate of infected insect cells has been used in an indirect ELISA to detect the presence of anti-NDV antibodies in a cohort of chicken sera. Data produced from these tests indicated a good correlation between ELISA titre and haemagglutination inhibition test data. The test was not affected by interference from background cellular proteins nor by cross-reactivity with non-NDV poultry pathogens. Additionally, the test did not generate false-positive readings.

背景与目标： 含有编码新城疫病毒（Ulster 2C株）的核衣壳（NP）蛋白的cDNA克隆的重组杆状病毒已被用于感染昆虫细胞（Spodoptera frugiperda）。观察到高水平的过表达NP蛋白，其占总细胞蛋白的高达40％，其随后被证明是抗原性的。源自被感染昆虫细胞的粗制可溶性裂解物的核蛋白已用于间接ELISA中，以检测一群鸡血清中抗NDV抗体的存在。这些测试产生的数据表明ELISA滴度和血凝抑制测试数据之间具有良好的相关性。该测试不受背景细胞蛋白的干扰，也不受与非NDV家禽病原体交叉反应的影响。此外，该测试未产生假阳性读数。

BACKGROUND & AIMS: The aims of the study were to compare glycohaemoglobin (HbA1c) values measured by DCA (a benchtop analyzer primarily designed for within-clinic rapid HbA1c determination) to a reference HbA1c method and home blood glucose monitoring, and to explore the possibility of an uniform expression of data. A total of 103 blood samples and the corresponding mean capillary glucose values (4.4 +/- 1.2 tests/day) of the preceding 2 months were collected from 34 insulin-dependent diabetic adults. We measured the correlations and agreements using the residual plots method and regression equations between HbA1c measured by DCA and high-pressure liquid chromatography (HPLC), and between DCA and capillary glucose values. A highly significant correlation (r2 = 0.85, P < 0.001) and an acceptable agreement (97% of values within 2 SD of the mean difference of 0.9% +/- 0.4%) was found between DCA and HPLC values. The regression equation calculated on the first half of the cases wasDCA (%) = 0.72 HPLC (%) +1.38. Of DCA values expressed in HPLC terms using this equation 87% fell within a clinically acceptable confidence interval when compared with measured HPLC data. A significant correlation (r2 = 0.40, P < 0.

01) was found between DCA and capillary glucose values, and the regression equation wasDCA (%) = 0.34 capillary glucose (mM) +4.44. Of glycaemic levels calculated from DCA values using this formula 82% fell within a clinically acceptable error range when compared with measured glycaemic values. We conclude that the three methods of assessment of diabetes control are well correlated and that it is possible, with a degree of precision acceptable for the clinical setting, to express all data in uniform units, e.g. mM of capillary glucose or percentage of HPLC-HbA1c, though a simple correspondence table based on our transfer equations may be clinically sufficient and more handy.

背景与目标： 该研究的目的是将DCA（主要用于临床内快速HbA1c测定的台式分析仪）测得的糖化血红蛋白（HbA1c）值与参考HbA1c方法和家庭血糖监测进行比较，并探讨均一表达的可能性数据的。从34位胰岛素依赖型糖尿病成年人中收集了总共103份血液样本，以及前2个月的相应平均毛细血管葡萄糖值（4.4±1.2测试/天）。我们使用残差图法和通过DCA和高压液相色谱（HPLC）测量的HbA1c之间以及DCA和毛细管葡萄糖值之间的回归方程式，测量了相关性和一致性。在DCA和HPLC值之间发现高度显着的相关性（r2 = 0.85，P <0.001）和可接受的一致性（97％的值在2 SD之内的平均差为0.9％/-0.4％）。在上半部分病例中计算出的回归方程为DCA（％）= 0.72 HPLC（％）1.38。与测量的HPLC数据相比，使用该公式以HPLC术语表示的DCA值中有87％处于临床可接受的置信区间内。发现DCA与毛细血管葡萄糖值之间存在显着相关性（r2 = 0.40，P <0> 01），回归方程为DCA（％）= 0.34毛细血管葡萄糖（mM）4.44。与测量的血糖值相比，使用该公式从DCA值计算出的血糖水平中，有82％属于临床可接受的误差范围。我们得出的结论是，三种评估糖尿病控制的方法之间具有很好的相关性，并且有可能以临床环境可接受的精确度，以统一单位表示所有数据，例如mM毛细血管葡萄糖或HPLC-HbA1c的百分比，尽管基于我们的转移方程式的简单对应表在临床上可能就足够了，而且更方便。

BACKGROUND & AIMS: :A sensitive sandwich enzyme immunoassay for human pulmonary surfactant protein D (SP-D) was developed and used to examine the blood SP-D levels of drowning victims. Human SP-D was purified from amniotic fluid by chromatographic methods, and an antibody against human SP-D was prepared. A polystyrene ball coated with anti-SP-D IgG was incubated with purified human SP-D, and then with anti-SP-D Fab'-peroxidase conjugate. Peroxidase activity bound to the polystyrene ball was assayed by fluorometry using 3-(4-hydroxyphenyl)propionic acid as the hydrogen donor. The detection limit of human SP-D was 5.2 pg per assay tube. Examination of cross-reactions of this sandwich enzyme immunoassay with proteins from other human organs showed it to be highly specific for lung, and Northern blot analysis detected specific SP-D mRNA expression only in lung. The SP-D concentration of normal human serum was 6.4+/-2.7 (mean+/-S.D.) ng ml(-1) (n=20). The recovery rates of 0.52 ng and 5.2 ng SP-D added to 5 microl normal human serum were 93.6+/-2.7% and 93.6+/-6.1%, respectively. Blood SP-D levels of victims from the saltwater drowning group (n=14) revealed higher concentrations (105.8+/-53.7 ng ml(-1)), while freshwater drowning victims (n=12) were estimated to be 74.1+/-43.9 ng ml(-1). The SP-D levels of 15 subjects who died of hemorrhage (n=5), heart failure (n=8), traumatic shock (n=1), and electrocution (n=1) were lower (22.0+/-8.5 ng ml(-1)), and those of asphyxia victims (n=10) were slightly higher (36.2+/-17.1 ng ml(-1)) than those of other causes of death, except for drowning. These results suggest that in drowning victims, SP-D flowed into the systemic circulation by physiological and physical mechanisms, and the differences of blood SP-D levels between saltwater drowning and freshwater drowning victims are presumed to be influenced by the type of agony and/or the length of survival time in water.

背景与目标： ：开发了一种针对人肺表面活性物质D（SP-D）的敏感三明治酶免疫测定法，并用于检查溺水受害者的血液SP-D水平。通过色谱法从羊水中纯化出人SP-D，并制备了针对人SP-D的抗体。将涂有抗SP-D IgG的聚苯乙烯球与纯化的人SP-D孵育，然后与抗SP-D Fab'-过氧化物酶偶联物孵育。使用3-（4-羟苯基）丙酸作为氢供体，通过荧光分析法测定了与聚苯乙烯球结合的过氧化物酶活性。每个分析管中人SP-D的检出限为5.2 pg。对该夹心酶免疫测定法与其他人体器官蛋白质的交叉反应的检查表明，它对肺具有高度特异性，Northern印迹分析仅在肺中检测到特定的SP-D mRNA表达。正常人血清的SP-D浓度为6.4 /-2.7（平均值/-S.D。）ng ml（-1）（n = 20）。添加到5微升正常人血清中的0.52 ng和5.2 ng SP-D的回收率分别为93.6 /-2.7%和93.6 /-6.1%。盐水淹没组（n = 14）受害者的血液SP-D水平显示较高的浓度（105.8 /-53.7 ng ml（-1）），而淡水淹没的受害者（n = 12）估计为74.1 /-43.9 ng ml（-1）。 15名死于出血（n = 5），心力衰竭（n = 8），外伤性休克（n = 1）和电击伤（n = 1）的受试者的SP-D水平较低（22.0 /-8.5 ng ml （-1）），窒息受害者（n = 10）的死亡人数（36.2 /-17.1 ng ml（-1））略高于其他死亡原因，但溺水除外。这些结果表明，在溺水受害者中，SP-D是通过生理和物理机制流入全身循环的，据推测，海水淹没受害者与淡水溺水受害者之间血液SP-D水平的差异受痛苦和//类型的影响。或在水中生存时间的长短。

BACKGROUND & AIMS: A noncompetitive enzyme immunoassay method (hetero-two-site enzyme immunoassay) for salmon calcitonin (SCT) and its usability for the pharmacokinetic study are described. The method in brief proceeds as followscentrifugal filtration through a polysaccharide membrane to remove plasma proteins, biotinylation, trapping onto an anti-SCT IgG-coated polystyrene ball, acid elution, coupling with affinity-purified anti-SCT Fab'-peroxidase conjugate, final trapping onto streptavidin-coated polystyrene balls, and measurement of peroxidase activity bound to the balls by fluorometry. The practical detection limit of SCT was 0.1 pg (30 amol)/assay and 2 pg/ml as the assay sample's concentration, which was at least fivefold lower than those previously reported by competitive radioimmunoassays. The application of this method has enabled us to 1) directly estimate the bioavailability of SCT dosed subcutaneously at the therapeutic levels (1.2 and 4.7 micrograms/kg) for its antiosteoporotic effect as compared to an intravenous dose (1.2 micrograms/kg) and 2) search for the relationship between blood level and the hypocalcemic activity of SCT. The pharmacokinetic parameters of subcutaneous SCT (1.2 and 4.

7 micrograms/kg) thus estimated were as followsthe area under the blood concentration-time curve (AUC) = 89 and 550 pg.hr/ml, and mean residence time (MRT) = 44 and 65 minutes, respectively, when the AUC for an intravenous SCT (1.2 micrograms/kg) = 160 pg.hr/ml and the MRT = 10 minutes.

背景与目标： 描述了鲑鱼降钙素（SCT）的非竞争性酶免疫测定方法（异位两点酶免疫测定）及其在药代动力学研究中的可用性。该方法的简要介绍如下：通过多糖膜离心过滤以除去血浆蛋白，进行生物素化，捕获到抗SCT IgG包被的聚苯乙烯球上，酸洗脱，与亲和纯化的抗SCT Fab'-过氧化物酶偶联物偶联，最后捕获到抗生蛋白链菌素包被的聚苯乙烯球上，并通过荧光法测量结合到该球上的过氧化物酶活性。 SCT的实际检出限为0.1 pg（30 amol）/测定，测定样品的浓度为2 pg / ml，比竞争性放射免疫测定所报道的浓度低至少五倍。这种方法的应用使我们能够（1）直接估计以治疗水平（1.2和4.7微克/ kg）皮下注射的SCT的抗骨质疏松作用的生物利用度，而静脉注射剂量（1.2微克/ kg）和2）寻找血液水平与SCT降钙活性之间的关系。如此估算的皮下SCT的药代动力学参数（1.2和4 7微克/ kg）如下：血液浓度-时间曲线下的面积（AUC）分别为89和550 pg.hr/ml，均值当静脉内SCT的AUC（1.2微克/千克）= 160 pg.hr/ml且MRT = 10分钟时，停留时间（MRT）分别为44分钟和65分钟。

BACKGROUND & AIMS: :Serotyping of Shiga toxin-producing Escherichia coli (STEC) has been contingent upon the availability of antisera. Here we describe a 7-plex microbead-based immunoassay to simultaneously serotype seven STECs (i.e., belonging to serogroups O26, O45, O103, O111, O121, O145, and O157) by the Luminex xMAP® technology. This technology presents many advantages: Its multiplexed format (up to 100 analytes) saves time, reagents, and test sample, and many regulatory agencies currently utilize this platform for other assays. In this study, a total of seventy-nine STEC strains belonging to the 7 different serogroups of interest were tested. These strains had been previously serotyped and their serogroup was confirmed by PCR. Except for one strain belonging to the O111 serogroup, nearly all strains (i.e., 98.7%; 78/79) were correctly identified on the Bio-Plex 100 instrument in less than 4h. This newly developed microbead-based immunoassay could be extended to include other STEC serogroups, virulence factors, and/or bacterial species.

背景与目标： ：产生志贺毒素的大肠杆菌（STEC）的血清分型取决于抗血清的可用性。在这里，我们描述了一种基于7重微珠的免疫测定方法，通过LuminexxMAP®技术同时对7个STEC（即O26，O45，O103，O111，O121，O145和O157血清群）进行血清分型。这项技术具有许多优点：它的多重格式（最多100种分析物）节省了时间，试剂和测试样品，并且许多监管机构目前都将该平台用于其他测定。在这项研究中，共测试了7种STEC感兴趣的血清型，共79种。这些菌株先前已进行血清分型，并通过PCR确证了它们的血清组。除了一个属于O111血清群的菌株外，几乎所有的菌株（即98.7％； 78/79）都可以在不到4小时的时间内在Bio-Plex 100仪器上正确识别。这种新开发的基于微珠的免疫测定法可以扩展到包括其他STEC血清群，毒力因子和/或细菌种类。

BACKGROUND & AIMS: :Several glycosylated macromolecules associated with normal and malignant pancreatic ductal cells have been described. We have generated a monoclonal antibody, LD-B1, by immunizing Balb/c mice with the postmicrosomal extract of fresh human pancreatic ductal carcinoma tissue and used it in this study to characterize the nature of the target antigen. The antigen detected by LD-B1 antibody was purified to homogeneity by affinity chromatography. Enzymatic and biochemical analysis showed it to be a nonsialylated glycoprotein that on Western blotting and immunoprecipitation analyses had an apparent molecular weight of 300-400 kDa. The mobility on gels was not affected by reducing or denaturing conditions. Competitive inhibition assays with various MoAbs and lectins indicated that the epitope recognized by LD-B1 antibody involves the carbohydrate sequence Gal beta 1----3Gal beta 1----3(or 4)GlcNAc beta 1----3Gal. Using a double determinant sandwich ELISA, elevated antigen levels were detected in the sera of 5 of 6 patients with pancreatic carcinoma, 0 of 3 patients with chronic pancreatitis, and 12 of 137 normal controls. These results suggest that patients with pancreatic carcinoma exhibit altered expression of a heavily glycosylated molecule related to a blood group precursor immunodeterminant.

背景与目标： ：已经描述了与正常和恶性胰腺导管细胞有关的几种糖基化大分子。我们已经用新鲜的人胰腺导管癌组织的微粒体提取物免疫Balb / c小鼠，从而产生了单克隆抗体LD-B1，并将其用于本研究中以表征靶抗原的性质。用LD-B1抗体检测的抗原通过亲和层析纯化至均质。酶和生化分析表明它是一种非唾液酸化的糖蛋白，经蛋白质印迹和免疫沉淀分析，其表观分子量为300-400 kDa。凝胶上的迁移率不受还原或变性条件的影响。各种MoAb和凝集素的竞争性抑制分析表明，LD-B1抗体识别的表位涉及碳水化合物序列Gal beta 1 ---- 3Gal beta 1 ---- 3（或4）GlcNAc beta 1 ---- 3Gal。使用双重确定性夹心ELISA，在6例胰腺癌患者中有5例血清，3例慢性胰腺炎患者中有0例和137例正常对照中的血清中检测到抗原水平升高。这些结果表明，胰腺癌患者表现出与血型前体免疫决定簇有关的重糖基化分子的表达改变。

BACKGROUND & AIMS: BACKGROUND:Giardia duodenalis is conventionally diagnosed in fecal samples using parasitological methods. However, sensitivity is poor when only a single sample is analyzed, due to intermittent excretion of cysts in feces. Alternatively, the serum antibodies to G. duodenalis can be used for parasite diagnosis and epidemiological studies to determine previous exposure. We compared the rate of G. duodenalis infection between serum anti-Giardia IgG and IgA antibodies and fecal examination in Brazilian children. METHODS:Fecal and serum samples were tested from 287 children at a clinical laboratory and from 187 children at daycare centers. Fecal samples were processed using conventional parasitological methods and coproantigen detection for Giardia diagnosis. Serum samples were tested using an in-house ELISA for detection of anti-Giardia IgG and IgA. RESULTS:G. duodenalis was found in 8.2% (N=39) of the 474 children analyzed. The sensitivity and specificity of ELISA were 80.0% and 90.0% for IgG and 80.0% and 83.3% for IgA, respectively. The total positivity rate of anti-Giardia IgG and IgA in the sera was 13.9% (N=66) and 23.6% (N=112). The agreement between the positivity of specific antibodies and the detection of G. duodenalis in feces was moderate for ELISA-IgG, kappa index (95% CI)=0.543 (0.422-0.664), and mild for ELISA-IgA, kappa index (95% CI)=0.283 (0.162-0.404). Among the children infected with other enteroparasites, 11.6% (N=10) and 24.4% (N=21) showed reactivity to anti-Giardia IgG and to IgA, respectively. This cross-reactivity was more frequent in samples from children infected with Endolimax nana and Entamoeba coli. CONCLUSIONS:The higher frequency of specific antibody reactivity compared with G. duodenalis diagnosis in feces could reflect continuous exposure of children to G. duodenalis infection, resulting in long-lasting immunological memory and/or cross-reactivity with other intestinal amoebas.

背景与目标： 背景：传统上使用寄生虫学方法在粪便样本中诊断十二指肠贾第鞭毛虫。但是，由于粪便中的囊肿是间歇性排泄的，因此仅分析单个样品时灵敏度就很差。或者，可将十二指肠球菌的血清抗体用于寄生虫诊断和流行病学研究，以确定先前的暴露水平。我们比较了巴西儿童血清抗贾第鞭毛虫IgG和IgA抗体与粪便检查之间的十二指肠球菌感染率。
方法：从临床实验室的287名儿童和日托中心的187名儿童中检测粪便和血清样本。使用常规的寄生虫学方法和粪便原抗原检测处理粪便样本，以进行贾第虫诊断。使用内部ELISA测试血清样品以检测抗贾第鞭毛虫IgG和IgA。
结果：G。在分析的474名儿童中，发现十二指肠的比例为8.2％（N = 39）。 ELISA的灵敏度和特异性对IgG分别为80.0％和90.0％，对IgA分别为80.0％和83.3％。血清中抗贾第鞭毛虫IgG和IgA的总阳性率为13.9％（N = 66）和23.6％（N = 112）。特异性抗体的阳性与粪便中十二指肠的检测之间的一致性对于ELISA-IgG，κ指数（95％CI）= 0.543（0.422-0.664）为中等，而对于ELISA-IgA，κ指数（95）为中等％CI）= 0.283（0.162-0.404）。在感染了其他肠寄生虫的儿童中，分别有11.6％（N = 10）和24.4％（N = 21）表现出对抗贾第鞭毛虫IgG和IgA的反应性。在被Endolimax nana和Entamoeba coli感染的儿童的样本中，这种交叉反应更为频繁。
结论：粪便中特异性抗体反应的频率高于十二指肠杆菌的诊断，可能反映儿童持续暴露于十二指肠杆菌感染，从而导致长期的免疫记忆和/或与其他肠变形虫的交叉反应。