慢性硬膜下血肿引流液检测NSE的应用价值

目的：观察慢性硬膜下血肿（CSDH）患者引流液的神经元特异性烯醇化酶（NSE）的浓度,及其与血清NSE浓度的关系。方法：收集CSDH患者经钻颅所产生的引流液,同时采集病人和正常对照者的血液,经处理后采用放免法测定NSE的浓度。结果：CSDH患者组的血清NSE高于正常对照组（P〈0.01）;CSDH患者引流液的NSE与自身血清NSE相关（r=0.917）,引流液NSE高于自身血清NSE（P〈0.01）。结论：测定CSDH患者引流术后的引流液NSE浓度变化可以作为疗效观察的指标之一。     

S-100B、NSE和MBP对急性脑出血预后的临床评估

目的:研究脑出血后血清S-100B蛋白、神经特异性烯醇化酶(NSE)和碱性髓鞘蛋白 (MBP)浓度在预后评估中的价值。方法:对40例急性脑出血住院病人在病后72 h内进行血清S- 100B、NSE和MBP浓度检测,并结合格拉斯哥昏迷量表(GOS)评分进行比较分析。结果:本组40例病人脑出血后血清S-100B、NSE和MBP浓度均显著高于正常对照组,而且病人中不同预后组之间S- 100B、NSE和MBP浓度存在显著差异。分别以脑出血后72 h血清S-100B浓度2．0μg／L,NSE浓度 30μg／ml和MBP浓度10 μg／ml为临界标准评估顶后,S-100B的特异度为91％,敏感度为72％;NSE 特异度为77％,敏感度为67％;MBP特异度为63％,敏感度为61％。结论:脑出血后血清S-100B蛋白、NSE和MBP浓度对评估脑出血病人预后具有较高的特异性和敏感性。     

肝移植患者围术期血浆S-100β和神经元特异性烯醇化酶的变化

目的:观察不同病种、不同Child-Turcotte-Pugh(CTP)评分和终末期肝病模型(MELD)评分的肝移植患者血浆S-100β和神经元特异性烯醇化酶(NSE)的变化。方法:选择行肝移植手术的30例患者,于麻醉后(T1)、术毕时(T2)及新肝24h(T3),经左侧颈内静脉逆行置管至颈静脉球部,取血样检测S-100β和NSE浓度;并根据患者的病种、CTP和MELD评分进行分组,比较各组间S-100β和NSE浓度的变化。结果:T2时重肝组S-100β和NSE浓度均高于肝硬化组,CTP分级C级患者T2时的S-100β浓度高于A级患者;CTP分级C级患者T1时的NSE浓度高于A级患者;3个时点≥18分组的S-100β浓度均高于11分组,T1和T2时≥18分组的NSE浓度高于11分组。结论:病情越重,肝功能越差的患者,其S-100β和NSE浓度越高,表明脑损伤的程度越重。     

新生儿窒息、缺氧缺血性脑损伤与神经元特异性烯醇化酶浓度的相关性探讨

目的探讨新生儿血清神经元特异性烯醇化酶(NSE)浓度变化与新生儿窒息、缺氧缺血性脑损伤(HIBD)发生率及严重程度的相关性。方法用免疫放射法对30例窒息儿生后1天、3天、5天及14天的血清进行NSE测定,100例正常新生儿生后3天血清进行NSE测定作为对照。结果两组生后3天血清NSE均数之间比较,窒息组较正常组高,差异有显著性(P(0.05)。窒息组病情好转NSE随之下降,生后第1天与第5天、14天的血清NSE两两比较差异有显著性(P(0.05);第3天与14天的血清NSE均数之间的比较差异有显著性(P(0.05);窒息儿中67%的NSE浓度高于正常组,而且窒息程度越重,NSE浓度越高,HIBD发生率越高。结论生后第3天血清NSE浓度窒息新生儿较正常新生儿高;NSE浓度变化与缺氧缺血性脑损伤的病程相符,与病情严重程度呈正相关;NSE与窒息发生率及窒息的严重程度有较密切     

分型测定脑梗死急性期患者血清NSE和S-100β的变化

目的:探讨脑梗死亚型急性期血清神经元特异性烯醇酶(NSE)和S-100 β蛋白(S-100 β)变化的临床意义。方法:59例急性脑梗死被分为全前循环梗死(TACI)、 部分前循环梗死(PACI)、腔隙性梗死(LACI)和后循环梗死(POCI),以酶联免疫吸附试验(ELISA)测定起病6 d内血清NSE和S-100 β浓度的变化, 同对照组比较。结果:TACI各时点的血清NSE浓度均高于对照组(P＜0.01),于第3 d达到高峰, PACI各时点的NSE浓度亦高于对照组(P＜0.05),于1 d 达高峰。TACI的血清S-100 β浓度与NSE同步升高;PACI的S-100 β浓度则于3 h 开始上升,1 d达高峰,6 h、1 d、3 d和6 d的浓度明显高于对照组(P＜0.05)。LACI、POCI的血清NSE和S-100 β无明显改变。结论:脑梗死亚型急性期(含超早期)血清NSE和S-100 β浓度变化不同,此或许有助于分型处理急性脑梗死及其病理生理研究。     

NSE和S-100蛋白在新生儿窒息中的临床应用

目的通过检测窒息新生儿血清中神经特异性烯醇化酶（neuron—specific enolase,NSE）和S-100蛋白含量的变化。建立对窒息患儿的早期预后评估技术指标。并探讨NSE和S一100蛋白浓度之间的相关关系。方法采用酶联免疫吸附试验双抗体夹心法检测,对156例窒息患儿与对照组20例健康新生儿第3天血清NSE和S-100蛋白含量进行检测,并追踪随访,于1岁时进行Ge sell测试。结果窒息新生儿血清NSE和S-100蛋白含量显著高于对照组（P〈0．05）;HIE组和无HIE组的血清NSE和S-100蛋白浓度比较有显著差异（P〈0．05）;预后良好的窒息新生儿其血清NSE和S-100浓度较预后不良者低（P〈0．05）;NSE和S-100蛋白相关系数为0．756,经检验具有统计学意义（P〈0．05）。结论窒息新生儿血清NSE和S-100蛋白浓度可联合作为窒息后脑损伤的生化学评估指标。     

血清NSE和S—100B蛋白在急性脑梗死患者预测和预后评估中的临床意义

现代影像学技术的发展为脑血管病诊断提供了很大的帮助，但对于临床因病情危重而不宜搬动的病人则受到很大限制，国内外研究结果表明，血清神经元特异性烯醇化酶（NSE）和S-100B蛋白浓度测定作为病理学标志蛋白以及中枢神经系统损伤时的定量指标，同其高度敏感性和特异性，正逐渐被临床所重视。NSE特征性位于神经元和神经内分泌细胞质中。S-100B蛋白是神经胶质细胞的标志性蛋白。本文应用电化学发光免疫分析技术和酶联免疫分析法（ELISA）分别测定急性脑梗死患者血清NSE和S-100B蛋白浓度，并探讨其在病情预测及预后中的临床意义。     

颅脑损伤患者血清S-100B和神经元特异性烯醇化酶的检测

目的探讨颅脑损伤患者血清S鄄100B和神经元特异性烯醇化酶(NSE)的变化及其临床意义。方法用双抗体夹心ELISA法检测30名健康人和108例颅脑损伤患者血清S鄄100B和NSE水平。结果108例颅脑损伤患者在伤后12h血清S鄄100B水平[(1.24±0.32)滋g/L]较对照组[(0.38±0.12)滋g/L]升高,重型、中型和轻型颅脑外伤患者的血清NSE浓度均明显高于对照组的NSE浓度(P<0.01);而不同组别颅脑外伤患者血清NSE水平差异亦有显著性(P<0.05)。结论血清S鄄100B和NSE检测可作为诊断早期脑损伤的指标之一。     

急性颅脑外伤患者血清NSE含量变化及临床意义

目的：探讨急性颅脑外伤患者血清神经元特异性烯醇化酶（Neuron-specificenolase,NSE）的含量及动态变化对颅脑损伤的诊断和评估伤情、观察疗效、预后判断的临床价值。方法：采用发光免疫分析,对60例急性颅脑外伤患者及62例正常人血清NSE含量进行同步检测。并于治疗前和治疗后3d、1周、2周、3周NSE的含量进行动态观察。对38例不同预后的急性颅脑外伤患者治疗前后的血清NSE浓度及GCS评分进行回顾分析。结果：急性颅脑外伤患者血清NSE含量显著增高,与正常对照组比较差异具有极显著意义（P〈0.001）,其NSE增高的程度,重型组〉中型组〉轻型组（P〈0.01）,与GCS评分密切负相关（r=-0.9140,P〈0.01）。动态观察显示56例不同程度的颅脑外伤患者治疗后血清NSE浓度随着治程的延续而逐渐下降,还发现急性颅脑外伤患者的血清NSE浓度如果高滴度持续增高,则预后不佳,反之则预后相对较好。而且NSE浓度下降越迅速,恢复正常的时间越早,其预后越佳。结论：血清NSE含量反映脑细胞损伤的程度,是一项灵敏的量化指标。进行NSE含量的检测不仅对脑损伤的诊断和评估伤情富有临床价值,而且对于疗效观察及预后的早期判断均有帮助。     

抗人脑神经元特异性烯醇化酶(NSE)单克隆抗体杂交瘤细胞株Ly-1的建立

神经元特异性烯醇化酶,简称NSE,是肺小细胞癌(SCLC)和儿童神经母细胞瘤高特异性的血清学标志物。NSE可用来鉴别SCLC和非小细胞癌(NSCLC),评价治疗反应,预报复发。做为一个十分有用的肿瘤标志,NSE已被用于临床病例分析。我们自人脑分离纯化出高纯度NSE抗原,免疫BA-     

Two ELISA methods for the measurement of airborne swine epithelial antigens

A new double antibody sandwich ELISA for measuring swine epithelial antigens in swineries was compared to an adaption of our previously developed ELISA inhibition method. The sandwich ELISA is more sensitive than the inhibition ELISA, and its reproducibility is better. The quantitative results from swinery samples of the sandwich ELISA are about ten times higher, but correlate well (rs 0.88, p = 0.004) with the results of ELISA inhibition. The progression in magnitude of the results in relation to the sample dilution was examined.     

一种检测可溶性白细胞介素2受体夹心法ELISA的建立

采用两种识别不同表位的小鼠抗人白细胞介素2受体(IL-2R)单克隆抗体(McAb),在国内首次建立了检测可溶性IL-2R(sIL-2R)的夹心法ELISA,并进行了初步应用。结果显示:本法特异性强、敏感性高、重复性好、操作简便,能检出正常人、某些患者血清中以及细胞培养上清中sIL-2R,可用于基础和临床免疫学研究。     

FITC与抗-FITC系统检测促黄体生成素的应用评价

目的 使用抗-异硫氰酸荧光素(FITC)固相包被板建立促黄体生成素(LH)的酶联免疫(ELISA)检测方法.方法用FITC及辣根过氧化物酶(HRP)分别标记两株抗-LH单克隆抗体,建立一步检测LH的ELISA检测方法,并与经典的双抗体夹心法进行了方法学对比评价.结果 FITC与抗-FITC系统检测LH,在2~50mIU/mL的校准曲线范围内相关系数0.9968,分析内精密度为7.6%,分析间精密度为7.02%,热稳定性下降18.5%,与双抗体夹心法相当,空白检测限为0.15mIU/mL,优于双抗体夹心法,钩状效应(HOOK效应)比双抗体夹心法差.与贝克曼检测结果回归方程Y=0.970X+0.614,相关系数r=0.975.结论 成功建立基于FITC与抗-FITC系统的ELISA检测方法,与双抗体夹心法相比,两种方法均能满足临床检测的需要.     

Preparation of thermo-responsive membranes. II

Two types of liquid crystal (LC)-immobilized membranes were prepared by a soaking method and sandwich method to control the permeation of indomethacin, as a model drug, in response to local and systemic fever. Monooxyethylene trimethylolpropane tristearate (MTTS) was used as a model LC because it has a gel-liquid crystal phase transition temperature near the body temperature, 39-40 degrees C in phosphate buffered saline (pH 7.4). Two porous polypropylene (PP) membranes were soaked into 20% MTTS chloroform solution in the soaking method, and two PP membranes were poured with the melted MTTS and pressed in the sandwich method. Thermo-response efficacy of the soaked membrane was dependent upon the content of MTTS in MTTS membrane, and the MTTS content above the void volume of PP membrane (38%) was needed for high efficacy. On the other hand, the sandwich membrane exhibited higher thermo-response efficacy than the soaked membrane, because more LC was embedded in the pores of sandwich membrane than that of the soaked membrane. The sandwich membrane permeation of indomethacin was sharply controlled by temperature changes between 32 and 38 degrees C.     

Comparison of an established antibody sandwich method with an inhibition method of Histoplasma capsulatum antigen detection

The Histoplasma antigen immunoassay utilizes an antibody sandwich method that provides a rapid and reliable means of diagnosing the more severe forms of histoplasmosis. Inhibition assays have been developed for antigen detection and offer at least one potential advantage, namely, reduced antibody requirements. We have developed an inhibition assay using the polyclonal antibody employed in our standard sandwich assay. Urine and serum specimens from patients with culture-proven histoplasmosis and controls were tested using both methods. The two methods had similar sensitivities for detection of antigen in urine (antibody sandwich = 92.5% versus inhibition = 87.5%, P = 0.500) and serum (82.5% versus 80.0%, P = 1. 000). With serum, the specificities of both methods were similar (antibody sandwich assay = 95.0% versus inhibition assay = 92.5%, P = 1.000), and with urine, the specificity of the antibody sandwich method was superior (97.5% versus 80.0%, P = 0.039). While the overall reproducibility of both methods was excellent (with urine, antibody sandwich assay intraclass correlation coefficient = 0.9975 and with serum = 0.9949; correlation coefficient of the inhibition assay with urine = 0.9736 and with serum = 0.9850), that of the inhibition method was only fair to poor for the controls: urine = -0. 0152, serum = 0.5595. Reproducibility was good for the controls using the sandwich method: urine = 0.7717, serum = 0.9470. Cross-reactivity was observed in specimens from patients infected with Blastomyces dermatitidis, Paracoccidioides brasiliensis, and Penicillium marneffei. In conclusion, the decreased specificity and inferior reproducibility with control specimens suggest that the inhibition assay has poorer precision toward the lower end of the detection range.     

A simplified cytokine immunoassay using magnetic polymer particles

Accurate detection of cytokines is essential for understanding their biological role in the immune system. Various methods to detect cytokines have been developed, including sandwich enzyme-linked immunosorbent assay (ELISA) and flow cytometry-based methods. All of the currently available methods have limitations, however. These limitations include time and extensive handling in standard sandwich ELISAs and the need for specialized equipment in flow cytometry-based assays. We have developed a magnetic polymer cytokine immunoassay and demonstrate that this assay is rapid and simple, needs less handling and offers better dynamic range, compared to standard sandwich ELISA. Furthermore, it does not require flow cytometry equipment, which is often used in microparticle-based polymer immunoassays. The magnetic polymer cytokine immunoassay described in this study is as sensitive as a standard sandwich ELISA. Because the method is not limited to the use of magnetic polymer particles, it is versatile and compatible with a number of different solid matrixes.     

多种单抗联合检测HIV抗原

目的 建立多种单抗联合早期检测HIV抗原的夹心ELISA方法.方法 以SAS盐析沉淀法和亲和层析法纯化抗HIV-1 p24、gp41、gp120及抗HIV-2 gp36的腹水型单克隆抗体(McAb),用高碘酸钠法将纯化的McAb以HRP进行标记.建立针对单个抗原的双抗体夹心ELISA法,对其灵敏度及特异性进行检测.将筛选得到的4株捕获McAb按比例混合作为捕获抗体,4株酶标McAb按比例混合作为检测抗体,建立多种单抗联合检测HIV抗原的夹心ELISA方法,检测混合HIV抗原.结果 按确定的最优反应条件建立的多种McAb联合夹心ELISA方法,检测到的最高稀释度的HIV混合抗原中各抗原的终浓度分别为:重组HIV-1 p24:0.625 pg/ml,gp41:6.25 ng/ml,gp120:6.25 ng/ml;HIV-2 gp36:9.25 ng/ml.结论 建立了具有高度敏感性的鸡尾酒式多种单抗联合检测HIV抗原的夹心ELISA法,为早期榆测HIV抗原提供了新的思路,为后续的研究奠定了一定基础.     

多种单抗联合检测HIV抗原

目的 建立多种单抗联合早期检测HIV抗原的夹心ELISA方法.方法 以SAS盐析沉淀法和亲和层析法纯化抗HIV-1 p24、gp41、gp120及抗HIV-2 gp36的腹水型单克隆抗体(McAb),用高碘酸钠法将纯化的McAb以HRP进行标记.建立针对单个抗原的双抗体夹心ELISA法,对其灵敏度及特异性进行检测.将筛选得到的4株捕获McAb按比例混合作为捕获抗体,4株酶标McAb按比例混合作为检测抗体,建立多种单抗联合检测HIV抗原的夹心ELISA方法,检测混合HIV抗原.结果 按确定的最优反应条件建立的多种McAb联合夹心ELISA方法,检测到的最高稀释度的HIV混合抗原中各抗原的终浓度分别为:重组HIV-1 p24:0.625 pg/ml,gp41:6.25 ng/ml,gp120:6.25 ng/ml;HIV-2 gp36:9.25 ng/ml.结论 建立了具有高度敏感性的鸡尾酒式多种单抗联合检测HIV抗原的夹心ELISA法,为早期榆测HIV抗原提供了新的思路,为后续的研究奠定了一定基础.     

检测肌糖蛋白C夹心ELISA方法的建立及初步应用

目的:以已经制备的抗肌糖蛋白C(TN-C)的单克隆抗体(mAb)为基础,建立能定量检测肌糖蛋白C浓度的夹心ELISA方法,并初步于临床血清标本检测.方法:将3株mAb(克隆号分别为:1A8、3H7和4D6)制备腹水后纯化,分别与辣根过氧化物酶交联后,两两配对,以重组肌糖蛋白C 蛋白为检测抗原,分析抗体之间最佳组合;利用建立的夹心ELISA方法检测收集的临床骨肉瘤患者和正常人血清标本.结果:包被1A8 mAb,HRP-4D6配对敏感性最高;骨肉瘤患者血清中肌糖蛋白C浓度明显高于正常人.结论:成功建立检测肌糖蛋白C的夹心ELISA方法,并利用其检测到肌糖蛋白C在骨肉瘤患者中的浓度异于正常人.     

Detection of Penicillinase in Milk by Sandwich ELISA Based Polyclonal and Monoclonal Antibody

A sandwich ELISA has been developed using polyclonal and monoclonal antibody for the determination of penicillinase in milk. For this purpose, specific polyclonal and monoclonal antibodies against penicillinase were generated and characterized. Using penicillinase standards prepared from 1–128 ng/mL, the method indicated that the detection limit of the sandwich ELISA, as measured in an ELISA plate reader, was as low as 0.86 ng/mL of penicillinase. For determine the accuracy, raw milk containing 2, 8, 32, and 64 ng/mL of penicillinase were tested by sandwich ELISA. Recoveries were from 93–97.5%, and the coefficient of variation [CV (%)] were from 5.55–8.38%. For interassay reproducibility, recoveries were from 89.5–95.1%, the coefficient of variation [CV (%)] were from 5.26–9.58%. This sandwich ELISA provides a useful screening method for quantitative detection of penicillinase in milk.