化学发光自显影检测人血中巨细胞病毒抗体

本文报道用化学发光自显影技术(Chemiluminescent photographic detection,CPD)与点免疫测定(Dot-immunobinding assay,DIA)结合检测了人血中巨细胞病毒(HCMV)抗体。从氧化剂、发光增强剂、缓冲液种类优选及浓度优选找出了化学发光废物液的最佳配方。以硝酸纤维素膜(NC)为载体,用x-光胶片记录化学发光,检测辣根过氧化物酶(HRP),检测限2×10~(-15)～10~(17)mol。用NC以来心法检测HCMV-Ab,与常规ELISA法对比,57例样品中,定性判断阴阳性彼此符合48例,符合率为84.2%(48/57),但CPD法所用酶标抗体浓度较ELISA法小千倍。     

锁阳中抗氧化活性成分的筛选研究

目的：筛选锁阳中的抗氧化活性成分。方法：采用CUPRAC法确定锁阳抗氧化活性部位。以Cu²⁺-新亚铜灵试剂为显影试剂的TLC-生物自显影技术结合硅胶柱层析筛选分离活性部位中的抗氧化活性物质,并采用¹H-NMR和¹³C-NMR技术确定其的化学结构。结果：确定乙酸乙酯部位为锁阳的抗氧化活性部位,筛分得到的抗氧化活性成分主要有原儿茶酸、儿茶素、没食子酸、儿茶素没食子酸酯。结论：以Cu²⁺-新亚铜灵试剂为显影试剂的TLC-生物自显影技术能够快速、有效地从锁阳活性部位中筛选出抗氧化活性成分。     

A—FP放射火箭自显影术方法的改进

A—FP放射火箭自显影术,用于早期诊断肝癌,具有专一性强、灵敏度与精确度较高的优点。但以往沿用的方法其流程平均需要三天左右才能完成。我们设计了一种改良的A—FP自显影术,一般一天即可完成,效果完全相同。改进的方法要点是在火箭电泳免疫沉淀放射自显影技术的基础上,各加用中速增感屏(或稀土增感屏)提高感光速度。一、材料: 1.上海生物制品研究所(或长春生物制品研究     

薄层色谱-生物自显影技术在活性物质筛选中的应用

薄层色谱-生物自显影技术(TLC-bioautography)将薄层色谱分离和生物活性测定相结合,是一种活性指导下的快速靶向追踪分离、筛选活性成分的方法。此法操作简单、耗时短、灵敏度和专属性高,常应用于抗细菌、抗真菌、抗氧化,以及对乙酰胆碱酯酶抑制剂、葡萄糖苷酶抑制剂等活性天然产物的筛选。笔者对薄层色谱-生物自显影技术在活性物质筛选方面的应用进行综述。     

化学发光技术在核酸分子杂交中的应用

化学发光的发现历史悠久,但把化学发光技术与核酸分子杂交结合起来,建立化学发光核酸分子杂交技术却是近几年的事。1983年Heller和Schneider首先将化学发光技术引入了核酸的分析,这是继建立发光免疫技术之后,化学发光技术应用的又一个新的领域。目前,有关这方面的报道还不多,本文试就化学发光核酸分子杂交技术及有关的应用作一简要的介绍。     

常用化学发光免疫分析技术及其特点

通过对常用化学发光免疫分析(Chemiluminescenctimmunoassay,CLIA)技术的详细介绍,全面归纳了几种化学发光免疫分析技术的基本原理,详细分析临床免疫检测中常见的化学发光免疫类型及特点,简要概述化学发光免疫分析技术的新进展.     

中药检验新技术在《中华人民共和国药典》的应用

为了促进现代仪器分析技术在中药检验中的应用,进一步提高和完善中药质量标准,综述2010年版《中华人民共和国药典》一部新增检验技术：DNA分子鉴定、中药指纹图谱、液相色谱 质谱联用技术、薄层色谱 生物自显影的应用概况.     

人参总皂甙对小鼠骨髓基质细胞RNA表达及其细胞因子诱生的影响

采用胶片法（细胞贴壁胶片生长法）及放射自显影等技术，较系统地观察了人参总甙（ＴＧＳ）对小鼠骨髓基质细胞ＲＮＡ表达的时相特点及对细胞因子ＩＬ－３、１１－６诱生的影响。结果表明．ＴＧＳ在体外对小鼠骨髓基质细胞ＲＮＡ的代谢有明显的促进效应．并且发现骨髓基质细胞实验上清中ＩＬ－３、ＩＬ－６因子的活性显著高于正常对照组。在此基础上。又利用放射自显影技术做为胶片法可行性的佐证．取得满意结果。     

内镜下逆行胰胆管造影介入治疗胆胰疾病的护理体会

逆行胰胆管造影(ERCP)是通过内镜活检孔道将导管自十二指肠乳头开口处插入,并经该导管逆行注入造影剂,使胆管及胰管在X线下显影的技术。1968年首先通过十二指肠镜成功地进行了乳头插管,并随着内镜的改进、插管技术及造影     

分析化学发光免疫测定技术在甲状腺肿瘤患者生化免疫检验中的应用价值

目的本次研究主要针对在甲状腺肿瘤患者的生化免疫检验当中采用化学发光免疫测定技术的效果展开分析。方法 2017年1月至2018年1月为本次研究时间所处区间,将区间内我院接诊的甲状腺肿瘤患者作为研究样本,研究样本数量为68例,患者均进行化学发光免疫测定技术。结果化学发光免疫测定技术的准确度、灵敏度与特异性接近于病理学准确诊断。结论将化学发光免疫测定技术应用在临床甲状腺肿瘤患者中的诊断价值比较高。     

A cyclic peptide–polymer probe for the detection of Clostridium botulinum neurotoxin serotype A

A Botulinum neurotoxin serotype A (BoNT/A) ELISA detection system was developed based upon an 11-mer cyclic peptide, termed C11-019, that was identified through peptide phage display technology. The assay employs a sandwich format using the C11-019 cyclic peptide attached to a PEMA (poly(ethylene maleic anhydride)) matrix as the capture phase and anti-BoNT/A polyclonal antibodies as the detection phase. Results reported demonstrate that the C11-019 peptide–polymer can specifically bind to BoNT/A with no cross-reactivity to other serotypes examined in assay buffers and a variety of body fluids and foodstuffs. When a highly sensitive chemiluminescent substrate was engaged, the detection of 1 pg/mL could be readily achieved within 3 h with a linear range of 0.1–1 ng/mL. These results demonstrate that an inexpensive peptide–polymer-based capture ELISA system can be used for rapid, sensitive and highly specific BoNT detection.     

Single-molecule detection and probe strategies for rapid and ultrasensitive genomic detection

This paper reviews the current state-of-the-art development of single-molecule detection (SMD)-based methods for ultrasensitive and specific analysis of genomic sequences. We first discuss several newly devised single fluorescent probe strategies that allow separation-free detection of low-abundance DNA sequences, such as quantum dot (QD)-mediated fluorescence resonance energy transfer (FRET) technology and dual-color fluorescence coincidence and colocalization analysis. Various schemes toward single DNA sizing and sequencing in solutions or on surfaces are also reviewed. In the end, we summarize the different microfluidic approaches developed for use with SMD to facilitate rapid, low-volume and quantitative analysis, such as electrokinetic and hydrodynamic single-molecule manipulation techniques.     

Flow-injection chemiluminescence determination of phentolamine based on its enhancing effect on the luminol-potassium ferricyanide system

It was found that the light emission produced by the oxidation of luminol by potassium ferricyanide in the basic medium was enhanced by phentolamine, a drug recently used to treatment of male and female sexual dysfunction. The optimum conditions for this chemiluminescent reaction were studied in detail by a flow-injection system. A new, simple and rapid method has been developed under the optimum conditions for determination of phentolamine. This method has the advantages of high sensitivity, good reproducibility and low detection limit. On the basis of investigation of chemiluminescent, fluorescent and UV spectra of phentolamine in basic solution containing potassium ferricyanide and luminol, a possible mechanism of this reaction was proposed. In the optimum conditions, CL intensities are proportional to concentrations of the phentolamine in the 0.01-1 microg/mL range. The limit of detection is 3.0 ng/mL for phentolamine. The method has been applied to the determination of phentolamine in the commercial preparations, synthetic samples and biological fluids with satisfactory results.     

三种检测方法检测抗dsDNA抗体的临床意义

目的探究将绿蝇短膜虫间接免疫荧光法、欧蒙酶免疫斑点法以及胶体金快速斑点法应用于检测抗双链DNA(dsDNA)抗体中的效果。方法120例系统性红斑狼疮患者,根据病情不同分为疾病活动组及疾病稳定组,每组60例;同时选取60例健康体检者作为对照组。分别采用绿蝇短膜虫间接免疫荧光法、欧蒙酶免疫斑点法以及胶体金快速斑点法对三组研究对象的抗双链DNA抗体进行检测,分析三种检测方式的检测阳性率。结果绿蝇短膜虫间接免疫荧光法对疾病活动组患者抗双链DNA抗体的检测阳性率为95.00%(57/60),对疾病稳定组患者的检测阳性率为71.67%(43/60),对对照组的检测阳性率为5.00%(3/60);欧蒙酶免疫斑点法对疾病活动组患者抗双链DNA抗体的检测阳性率为76.67%(46/60),对疾病稳定组患者的检测阳性率为61.67%(37/60),对对照组的检测阳性率为10.00%(6/60);胶体金快速斑点法对疾病活动组患者抗双链DNA抗体的检测阳性率为80.00%(48/60),对疾病稳定组患者的检测阳性率为65.00%(39/60),对对照组的检测阳性率为8.33%(5/60)。三种检测方法对疾病活动组患者抗双链DNA抗体的检测阳性率高于疾病稳定组、对照组,疾病稳定组高于对照组,差异具有统计学意义(P<0.05);绿蝇短膜虫间接免疫荧光法对系统性红斑狼疮患者抗双链DNA抗体的检测阳性率为83.33%(100/120),均高于欧蒙酶免疫斑点法的69.17%(83/120)、胶体金快速斑点法的72.50%(87/120),差异具有统计学意义(P<0.05)。结论三种诊断方式对于抗双链DNA抗体均具有较高的诊断阳性率,并且在对患者的各个时期进行诊断时,产生的诊断效果较为良好,而三种诊断方式应用于患者的诊断中,可以选择联合诊断,这样有助于避免误诊或者漏诊。     

A sensitive chemiluminescent enzyme immunoassay for the bioanalysis of carboxyl-terminal B-chain analogues of human insulin.

Quantification of analogues of human insulin in biological matrices is complicated by differences in their immunoreactivity and the presence of both the analogue and endogenous concentrations of insulin in test samples. To facilitate pharmacokinetic comparisons of carboxyl-terminal B-chain analogues of human insulin, we undertook development of a sensitive ELISA. The ELISA detection method was optimized systematically to permit routine analysis of 10-microl serum samples. Accordingly, a noncompetitive 'sandwich' chemiluminescent ELISA was validated for the quantification of carboxyl-terminal B-chain insulin analogues in human serum over a concentration range from 5 to 3125 pM. The mean bias (RE%) within the validated range varied from -10.3 to 4.3%, with an intermediate precision (inter-assay CV%) from 4.2 to 11.5%. The two-sided 90% expectation tolerance interval for total measurement error was within +/-25% of the nominal concentration for all levels of validation samples. Insulin lispro, human insulin, proinsulin, despentapeptide insulin (DPI) and porcine insulin displayed comparable crossreactivity in the ELISA. Potential utility of the new assay for insulin bioanalysis in nonhuman species was investigated by assessing the pharmacokinetic profile of DPI in rats following administration of a single subcutaneous dose. The sensitive chemiluminescent detection method is simple to perform and should be readily adaptable for ELISAs of other therapeutic proteins.     

Sensitive and multiplexed detection of proteomic antigens via quantum dot aggregation

A rapid, single-step, solution-phase method for quantifying multiple proteomic biomarkers is described. Nanoscale quantum dot–antibody conjugates self-assemble into microscale aggregates in the presence of a specific antigen through antibody-antigen molecular recognition. These aggregates are easily discriminated from the individual components by flow cytometry. Quantum dot (QD) aggregates can be quantified and correlated to the antigen concentration. Two QD populations with distinct emission spectra are used for detecting two proteomics antigens in a single reaction volume. Multiplexed detection of vascular endothelial growth factor A and angiopoietin-2 is demonstrated at the physiologically relevant, picomolar concentration range. Nonmultiplexed detection of the antigens is also demonstrated, with a femtomolar sensitivity limit. This technique may be optimized for low-cost early detection and frequent screening of cancers and other diseases as well as detection of the biological response to therapy.From the Clinical EditorIn this paper a rapid, single-step, solution-phase method is described with multiplexed detection at physiological relevant picomolar concentration range and nonmultiplexed detection with a femtomolar sensitivity limit. The technique may enable low-cost early detection of cancers and other diseases as well as detection of the biological response to therapy.     

高出检测限浓度的肿瘤标志物的准确值检测

目的为得到一个高出检测限浓度肿瘤标志物的具体值,而选择一种更好的稀释液对样本进行稀释,以满足临床对疗效观察的需求。方法分别采用三种不同的稀释剂：①美国雅培AXSYM全自动化学发光仪配套的第4号缓冲液;②生理盐水;③肿瘤标志物的阴性血清（每种被测肿瘤正常值低90%以下的血清）,对美国雅培AXSYM全自动化学发光仪所测的高浓度肿瘤标志物CEA,CA125,T-PSA,进行稀释,与仪器自动稀释的结果和在仪器检测限内检测结果进行比较。结果手工稀释的结果与机器自动稀释的结果比较,CA125：4号缓冲液稀释与仪器自动稀释结果无显著差异,而用生理盐水和用阴性血清稀释与仪器自动稀释有统计学差异;CEA和T-PSA：三种稀释液与仪器自动稀释均无显著性差异。结论美国雅培AXSYM全自动化学发光仪配套的4号缓冲液,对超高浓度即仪器已经不能自动稀释的那部分肿瘤标志物的血清稀释的结果最好。CA125阴性血清由于基础值高,则不能用来对CA125进行稀释,但可以对CEA和T-PSA进行稀释,但是结果不如4号缓冲液和生理盐水准确。     

血清铁蛋白测定对老年恶性肿瘤贫血的鉴别诊断价值

目的　探讨血清铁蛋白 (SF)测定在老年恶性肿瘤贫血病因分类中的鉴别诊断价值。方法　以化学发光法测定恶性肿瘤贫血病人及对照组血清铁蛋白水平。结果　老年恶性肿瘤并贫血病人与对照组之间血清铁蛋白水平差异有非常显著意义 (P <0 0 1)。结论　血清铁蛋白测定对老年恶性肿瘤贫血鉴别诊断敏感性高 ,可作为老年贫血病因分类的筛查试验     

Chemiluminescent enzyme immunoassays: a review of bioanalytical applications

This review gives an overview of the most recent and innovative developments in the field of chemiluminescent immunoassays through carefully selected examples. First, assays using microtiter plates for high-throughput or multiplexed assays aiming to achieve more complex assays through the multiplication of parameters per wells will be described. Systems will then be presented that have been recently developed, motivated by integration and miniaturization of existing immunoassays in more complex experimental setups. Finally, enhanced-performance chemiluminescent biochips, based on chemiluminescent reaction intensification, will be introduced.