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。

BACKGROUND & AIMS: :The usual specimens submitted by a medical examiner for toxicological analysis include blood, urine, bile, vitreous humor, stomach contents, and solid-organ tissue. The detection of drugs in these specimens typically involves a combination of techniques including colorimetry, immunoassay, and gas chromatography. Although many laboratories rely principally on urine for the detection of drugs of abuse by immunoassay, these assays may be applied to other specimen types. An evaluation of Microgenics Corporation's cloned enzyme donor immunoassay (CEDIA) was conducted in order to evaluate its use in the detection of cocaine/cocaine metabolites in vitreous humor specimens. During a 14-month period, 392 vitreous humor specimens were analyzed by the CEDIA DAU Cocaine assay. Instrument parameters were set according to published manufacturer's guidelines. All presumptive positive immunoassay results prompted confirmatory testing and quantitation by gas chromatography-mass spectrometry (GC-MS) of other specimens including blood. Vitreous humor specimens were not tested by GC-MS. Using a approximately 100-ng/mL cutoff, the CEDIA assay produced 23 presumptive positive results, 22 of which were confirmed by GC-MS. The only specimen which could not be confirmed, elicited an immunoassay screen value near the cutoff limit. Routine analysis of blood, urine, bile, and/or bladder wash specimens by gas chromatography-nitrogen phosphorus detection revealed the presence of cocaine/cocaine metabolites in only 7 (31.8%) of the 22 confirmed cases. The concentration ranges of cocaine and benzoylecgonine in the blood specimens were none detected to 337 ng/mL and 17 to 8598 ng/mL, respectively. Cocaethylene was not detected in these cases. Analysis of vitreous humor specimens by CEDIA improved the detection rate of cocaine/cocaine metabolites by 0.7% in the cases submitted to our laboratory during the 14-month period.

背景与目标： ：由体检医师提交的用于毒理学分析的常规标本包括血液，尿液，胆汁，玻璃体液，胃内容物和实体器官组织。这些标本中的药物检测通常涉及多种技术的组合，包括比色法，免疫测定和气相色谱法。尽管许多实验室主要依靠尿液通过免疫测定法检测滥用药物，但这些测定法也可以应用于其他类型的标本。为了评估其在玻璃体液标本中可卡因/可卡因代谢产物检测中的用途，对Microgenics公司的克隆酶供体免疫测定法（CEDIA）进行了评估。在14个月内，通过CEDIA DAU可卡因分析法分析了392具玻璃体液标本。仪器参数是根据已发布的制造商指南进行设置的。所有推定的阳性免疫测定结果均提示通过气相色谱-质谱（GC-MS）对包括血液在内的其他标本进行确认性测试和定量。玻璃体液标本未经GC-MS检测。使用大约100 ng / mL的临界值，CEDIA分析产生了23个推测阳性结果，其中22个通过GC-MS确认。唯一无法确认的标本在临界值附近引发了免疫分析筛选值。通过气相色谱-氮磷检测对血液，尿液，胆汁和/或膀胱洗涤标本进行常规分析，结果表明，在22例确诊病例中，仅7例（31.8％）存在可卡因/可卡因代谢产物。没有检测到血液样本中可卡因和苯甲酰芽子碱的浓度范围分别为337 ng / mL和17至8598 ng / mL。在这些情况下未检测到可口乙烯。通过CEDIA对玻璃体液标本进行分析，在14个月内提交给我们实验室的病例中，可卡因/可卡因代谢物的检出率提高了0.7％。

BACKGROUND & AIMS: :Progastrin-releasing peptide (proGRP) is a precursor of gastrin-releasing peptide, a hormone which is secreted from neuroendocrine cells. It has been shown to be a useful serum marker for small cell lung cancer. We raised monoclonal antibodies (MAbs) against proGRP with the primary aim of establishing a sensitive immunoassay. Immunization was performed with recombinant proGRP (amino acids 31-98) conjugated to thyroglobulin or with a DNP-modified peptide. Seven of the MAbs recognizing both recombinant and cell line-derived peptide were characterized and epitope-mapped. Based on cross-inhibition studies the antibodies could be categorized into three main groups. The molecular epitope assignment was studied by using phages displaying proGRP peptides, random peptide libraries displayed on phage and by pepscan analysis utilizing 10-mer biotinylated peptides. Two of the MAbs (E146, E172) bound to a defined region on the N-terminal part of proGRP(31-98), three recognized conformational-dependent epitopes in the middle of the peptide (E179, E180, E181) and two bound to the C-terminal part (E149, E168). Consensus sequences were obtained for MAbs E146, E149 and E168. The binding kinetics of the MAbs was determined by surface plasmon resonance, and a time-resolved immunofluorometric assay was established.

背景与目标： ：胃泌素释放肽（proGRP）是胃泌素释放肽的前体，胃泌素释放肽是神经内分泌细胞分泌的一种激素。已经显示它是用于小细胞肺癌的有用的血清标志物。我们提出了针对proGRP的单克隆抗体（MAb），其主要目的是建立敏感的免疫测定。用与甲状腺球蛋白缀合的重组proGRP（氨基酸31-98）或DNP修饰的肽进行免疫。识别了重组和细胞系衍生肽的7种单克隆抗体，并进行了表位映射。基于交叉抑制研究，抗体可分为三大类。通过使用展示proGRP肽的噬菌体，展示在噬菌体上的随机肽文库以及利用10-mer生物素化肽进行的pepscan分析，研究了分子表位的分配。两个单克隆抗体（E146，E172）结合到proGRP（31-98）N末端部分的定义区域，肽中间的三个公认的构象依赖性表位（E179，E180，E181）和两个结合连接到C端部分（E149，E168）。获得了针对单克隆抗体E146，E149和E168的共识序列。通过表面等离振子共振测定MAb的结合动力学，并建立了时间分辨的免疫荧光测定法。

BACKGROUND & AIMS: :The purpose of this study was to determine whether increased serum soluble fms-like tyrosine kinase-1 (sFlt-1) and decreased placental growth factor (PlGF) levels in pre-eclampsia are related to the clinical features and laboratory parameters of the patients, including markers of inflammation, endothelial activation and injury, oxidative stress and trophoblast debris. A total of 54 pre-eclamptic patients, 58 healthy pregnant and 52 healthy non-pregnant women were involved in this case-control study. Serum sFlt-1 and PlGF levels were measured by electrochemiluminescence immunoassay. Serum levels of sFlt-1 and PlGF were significantly higher in pre-eclamptic patients and healthy pregnant women than in healthy non-pregnant women. In addition, pre-eclamptic patients had significantly higher sFlt-1 levels and significantly lower PlGF concentrations compared with healthy pregnant women. According to the subgroup analyses, sFlt-1 levels were significantly higher in severely pre-eclamptic patients than in those with mild pre-eclampsia, whereas pre-eclamptic patients with fetal growth restriction or preterm onset of the disease had significantly lower PlGF concentrations compared with those without intrauterine growth restriction or with a disease onset at term. In the pre-eclamptic group, there were significant positive correlations between serum sFlt-1 levels and systolic and diastolic blood pressure, serum levels of blood urea nitrogen and creatinine, as well as plasma levels of von Willebrand factor antigen, fibronectin and cell-free fetal DNA. Furthermore, serum PlGF concentrations of pre-eclamptic patients showed significant positive correlations with gestational age at disease onset and delivery, as well as with fetal birth weight, and significant inverse correlations with levels of blood urea nitrogen, creatinine and fibronectin. In conclusion, increased serum sFlt-1 and decreased PlGF levels are associated with blood pressure, renal and endothelial dysfunction, trophoblast deportation, as well as with a shorter duration of pregnancy, fetal growth restriction, the severity and preterm onset of the disease in pre-eclampsia. These findings indicate the central role of an angiogenic imbalance in the pathogenesis of this pregnancy-specific disorder.

背景与目标： ：本研究的目的是确定先兆子痫患者血清可溶性fms样酪氨酸激酶1（sFlt-1）升高和胎盘生长因子（PlGF）降低是否与患者的临床特征和实验室参数有关包括炎症，内皮细胞活化和损伤，氧化应激和滋养细胞碎片的标志物。该病例对照研究共涉及54名先兆子痫患者，58名健康孕妇和52名健康非孕妇。通过电化学发光免疫测定法测量血清sFlt-1和PlGF水平。子痫前期患者和健康孕妇的血清sFlt-1和PlGF水平显着高于健康非孕妇。此外，与健康孕妇相比，先兆子痫患者的sFlt-1水平明显升高，而PlGF浓度则明显降低。根据亚组分析，严重先兆子痫患者的sFlt-1水平显着高于轻度先兆子痫的患者，而有胎儿生长受限或疾病早发的先兆子痫患者的PlGF浓度则明显低于轻度先兆子痫的患者。没有子宫内生长受限或足月病发作的人。在子痫前期组中，血清sFlt-1水平与收缩压和舒张压，血清尿素氮和肌酐水平以及血浆von Willebrand因子抗原，纤连蛋白和无细胞血浆水平之间存在显着正相关。胎儿DNA。此外，先兆子痫患者的血清PlGF浓度与疾病发作和分娩时的胎龄以及胎儿出生体重呈显着正相关，与血尿素氮，肌酐和纤连蛋白的水平呈显着负相关。总之，血清sFlt-1升高和PlGF水平降低与血压，肾脏和内皮功能障碍，滋养细胞驱逐出境以及妊娠持续时间短，胎儿生长受限，疾病的严重性和早发有关-子痫。这些发现表明在这种妊娠特异性疾病的发病机理中血管生成失衡的核心作用。

BACKGROUND & AIMS: :A solid phase fluorescent immunoassay using polyacrylamide beads coated with rabbit anti-porcine parvovirus antibodies has been developed and utilized in the diagnosis of porcine parvovirus infection. The antibody-coated beads (immunobeads) were used both to detect virus in mummified fetal tissues and to demonstrate specific antibodies in serum and ovarian follicular fluid. The immunobeads assay (IBA) was as sensitive as ELISA but more sensitive than virus isolation using tissue culture and haemagglutination tests. Both mummified and normal fetuses were obtained after experimental infection of SPF pregnant gilts. Using immunobeads, porcine parvovirus antigen was found in all mummified fetuses, but was found in only 1 out of 17 normal fetuses.

背景与目标： ：已经开发了一种使用涂有兔抗猪细小病毒抗体的聚丙烯酰胺珠的固相荧光免疫测定方法，并将其用于诊断猪细小病毒感染。抗体包被的珠子（免疫脂质体）既可用于检测木乃伊胎儿组织中的病毒，又可用于证明血清和卵巢卵泡液中的特异性抗体。免疫珠测定（IBA）与ELISA一样灵敏，但比使用组织培养和血凝试验分离病毒更灵敏。在对SPF怀孕的小母猪进行实验性感染后，可以同时获得木乃伊和正常胎儿。使用免疫珠，在所有木乃伊胎儿中均发现了猪细小病毒抗原，但在17例正常胎儿中仅发现1例。

BACKGROUND & AIMS: :Background. The prognostic significance of c-erb B-2 in breast cancer remains controversial. The aim of this study was to determine the practical prognostic significance of c-erb B-2 protein status in breast cancer extracts, using an enzyme immunoassay. Methods. An enzyme immunoassay was used to measure levels of c-erb B-2 protein prospectively in 360 patients with breast cancer, using cytosol fractions prepared for steroid receptor assay. The status of c-erb B-2 protein was assessed using a cut-off value for positivity of 18 ng/mg protein. Univariate and multivariate analyses were performed. To evaluate the prognostic significance of c-erb B-2 protein status. Results. Levels of c-erb B-2 protein in tumor tissue extract ranged from 0 to 213.0 ng/mg protein (mean, 15.5 ng/mg protein). In 52 tumors (14.4 %) more than 18.0 ng/mg protein was detected, and these tumors were regarded as c-erb B-2 protein-positive. Correlations were found between c-erb B-2 protein positivity and large tumor size (>3 cm; P = 0.0095), higher histological grade (P < 0.0001), estrogen receptor negativity (P < 0.0001), and progesterone receptor negativity (P < 0.0001). There was also a marginally significant correlation between c-erb B-2 protein positivity and lymph node positivity. Multivariate analysis showed that c-erb B-2 protein status was a significant independent prognostic factor for disease-free survival, being strongly significant in patients with positive lymph nodes. Conclusion. c-erb B-2-positive breast cancers are biologically more aggressive and c-erb B-2 protein status could be a candidate as a prognostic factor for patients with breast cancer, being particularly valuable in patients with positive lymph nodes.

背景与目标： ：背景。 c-erb B-2在乳腺癌中的预后意义仍然存在争议。这项研究的目的是使用酶免疫测定法确定乳腺癌提取物中c-erb B-2蛋白状态的实际预后意义。方法。使用为类固醇受体测定准备的细胞溶质，采用酶免疫测定法在360例乳腺癌患者中前瞻性地测量c-erb B-2蛋白的水平。使用18 ng / mg蛋白质阳性的临界值评估c-erb B-2蛋白质的状态。进行了单因素和多因素分析。评估c-erb B-2蛋白状态的预后意义。结果。肿瘤组织提取物中c-erb B-2蛋白的水平范围为0至213.0 ng / mg蛋白（平均值为15.5 ng / mg蛋白）。在52个肿瘤（14.4％）中，检测到的蛋白含量超过18.0 ng / mg，这些肿瘤被认为是c-erb B-2蛋白阳性。发现c-erb B-2蛋白阳性与大肿瘤尺寸（> 3 cm; P = 0.0095），组织学等级较高（P <0.0001），雌激素受体阴性（P <0.0001）和孕激素受体阴性（P <0.0001）。 c-erb B-2蛋白阳性与淋巴结阳性之间也存在微弱的相关性。多变量分析表明，c-erb B-2蛋白状态是无病生存的重要独立预后因素，在淋巴结阳性患者中非常重要。结论。 c-erb B-2阳性乳腺癌在生物学上更具侵略性，c-erb B-2蛋白状态可能是乳腺癌患者的预后因素，在淋巴结阳性患者中尤其有价值。

BACKGROUND & AIMS: :A specific and precise double antibody immunoassay for human plasma apolipoprotein B (apoB) was developed and applied in normolipidemic and hyperlipidemic subjects. The intra-assay coefficient of variation was ca. 9%. The distributions of total apoB and low density lipoprotein (LDL) apoB in a randomly selected, healthy, fasting population (n = 349) was slightly skewed with a mean total apoB of 81 mg/100 ml and LDL apoB of 72 mg/100 ml. The 90th percentile cutoffs for total apoB and LDL apoB were 106 and 97 mg/100 ml, respectively. Regarding total apoB, women showed a statistically significant increase (r = 0.463, p less than 0.001) with age (30-65) and an average annual increment of plasma apoB of 1.1 mg/100 ml. In contrast, men showed only a slight increase of apoB from the 4th to 5th decade, with an average annual increment of 0.7 mg/100 ml (r = 0.201, 0.02 less than p less than 0.05). Similarly, regarding LDL apoB, women showed an increase of 1.0 mg/100 ml/year from the 4th to 7th decade (r = 0.501, p less than 0.001), whereas men's LDL apoB did not increase significantly with age (r = 0.114, 0.2 less than p less than 0.3, for ages 30-49). Six of ten normal young subjects showed essentially no physiological variation in fasting apoB levels over a 10-wk period, whereas four had a variation of ca. 5% or less. LDL apoB represented ca. 90% of the total apoB in normolipidemic and type II plasma samples (86% in type IV samples) but only 68% in type III plasmas (n = 7). The ratios of LDL cholesterol-LDL apoB were similar for the random and hyperlipoproteinemic groups, ranging from a high of 1.8 for type IIa to a low of 1.5 for type IV. The ratio of cholesterol to apoB was significantly elevated (p less than 0.002) in the d less than 1.006 fraction of the type III plasma samples compared to the random and type II groups.

背景与目标： ：开发了一种针对人血浆载脂蛋白B（apoB）的特异性和精确的双抗体免疫测定方法，并将其应用于正常血脂和高血脂受试者中。批内变异系数为。 9％。随机选择的健康禁食人群（n = 349）中总载脂蛋白和低密度脂蛋白载脂蛋白的分布略有偏差，平均总载脂蛋白为81 mg / 100 ml，低密度脂蛋白载脂蛋白为72 mg / 100 ml 。总apoB和LDL apoB的第90个百分位数分别是106和97 mg / 100 ml。关于总载脂蛋白B，女性随着年龄的增长（30-65岁）显示出统计学上的显着增加（r = 0.463，p小于0.001），血浆载脂蛋白B的年均增加量为1.1 mg / 100 ml。相反，从第4至第5个十年，男性仅显示apoB略有增加，平均每年增加0.7 mg / 100 ml（r = 0.201，比p小于0.05小0.02）。同样，关于LDL apoB，从第4到第7个十年，女性显示增加了1.0 mg / 100 ml /年（r = 0.501，p小于0.001），而男性的LDL apoB没有随着年龄的增长而显着增加（r = 0.114，对于30-49岁的儿童，0.2小于p小于0.3）。十名正常青年受试者中有六名在10周内基本没有空腹apoB水平的生理变化，而四名受试者的ca值则有变化。 5％以下。 LDL apoB代表约。在正常血脂和II型血浆样本中，总apoB占90％（IV型样本为86％），但在III型血浆中仅为68％（n = 7）。对于随机组和高脂蛋白血症组，LDL胆固醇-LDL apoB的比率相似，范围从IIa型的高1.8至IV型的低1.5。与随机和II型组相比，在III型血浆样品中d小于1.006的d中，胆固醇与apoB的比率显着提高（p小于0.002）。