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1.
微板式化学发光酶免疫分析法临床测定人血清中孕酮   总被引:2,自引:0,他引:2  
将抗兔IgG(即二抗)物理吸附于聚苯乙烯微孔板上作为通用固相,通过免疫反应制备固相抗体。采用辣根过氧化物酶催化鲁米诺-过氧化氢化学发光体系,建立了一种高通量、简便、快速的化学发光酶免疫分析方法用于临床测定人血清中的孕酮。对各种影响因素如免疫试剂的稀释度、发光底物选择、发光反应时间及温育条件等进行了考察和优化,最终选定的实验条件:孕酮抗体和HRP标记物的最佳稀释度分别为1∶100000和1∶15000;选用Ⅱ号发光底物,发光反应10min后测定;37℃水浴条件下温育1h。对建立的方法进行了评价。该方法的灵敏度为0.08μg/L;批内和批间相对标准偏差均在15%之内;低、中、高3个不同浓度值样品的平均回收率分别为101%、101%和94.4%(在87.8%~108%之间)。使用本方法和经典的放射免疫法同时对36份人血清样品进行测定,结果显示,本方法与放射免疫分析方法相关性良好,其相关系数为0.9502。  相似文献   

2.
虾肉中呋喃它酮代谢物化学发光酶免疫分析方法的建立   总被引:1,自引:0,他引:1  
采用对碘苯酚增强的HRP-鲁米诺-H2O2化学发光体系,建立了虾肉中呋喃它酮代谢物5-吗啉甲基-3-氨基-2-恶唑烷基酮(AMOZ)残留的间接竞争化学发光酶免疫分析(icCLEIA)检测方法。检测所用抗体是基因重组的抗AMOZ衍生物单链抗体。优化的icCLEIA最佳工作条件为:AMOZA-OVA包被浓度62.5μg/L,抗AMOZ衍生物单链抗体最佳稀释度为1∶10,竞争免疫反应时间45 min,HRP酶标记羊抗鼠抗体最佳稀释度为1∶10000,孵育时间50 min。本方法的IC50为1.38μg/L;灵敏度为0.09μg/L;线性范围为0.26~9.08μg/L(IC20~IC80);批内和批间相对标准偏差均小于15%;抗AMOZ衍生物单链抗体与其它硝基呋喃类抗生素及其代谢物均没有交叉反应,特异性良好。4个不同添加量的AMOZ加标样品的平均回收率分别为72.2%,73.4%,72.6%和78.6%。与HPLC-MS/MS法测定值进行比较发现,两种方法相关性良好(R2=0.9997)。本方法可用于水产品中AMOZ残留的快速检测。  相似文献   

3.
增强化学发光酶免疫法对猪肉中盐酸克伦特罗的检测   总被引:4,自引:0,他引:4  
建立了猪肉中克伦特罗(CLB)的直接竞争化学发光酶免疫检测(dc-CLEIA)方法。通过优化离子强度、pH值确立了化学发光酶免疫法的标准曲线,优化后dc-CLEIA的检出限可达0.02μg/L。猪肉中的盐酸克伦特罗用高氯酸提取、SPE净化后绘制基质曲线,基质曲线与标准曲线吻合,说明基质影响基本消除。测定0.10、1.0、5.0μg/kg 3个水平的添加回收率,平均回收率为83%~98%,批内与批间的相对标准偏差均小于15%。进一步研究了dc-CLEIA与HPLC两种方法测定猪肉样品的相关性,结果显示两者测定结果相关性良好,r=0.964 7,说明所建立的直接化学发光酶免疫方法可用于实际样品的检测,结果准确可靠。  相似文献   

4.
水体及人体液中的雌二醇的高灵敏度化学发光酶免疫分析   总被引:1,自引:0,他引:1  
采用高灵敏度的化学发光酶免疫分析方法(CLEIA)研究测定了环境水体及人体尿液中17β-雌二醇(17β-estradiol)的含量。以FITC-anti-FITC体系为固相,生物素-亲和素体系作为信号放大体系,高灵敏度的3-(2′-螺旋金刚烷)-4-甲氧基-4-(3″-羟基)苯1,2-二氧杂环丁烷磷酸钠(AMPPD)-碱性磷酸酶(ALP)化学发光反应作为信号检测体系。优化分析条件后,测量的线性范围为2.5~2000ng/L;检出限为1.1ng/L;批内、批间分析误差均在10%以内,稀释回收率和加标回收率均在92%~112%之间。测得环境水样中雌二醇的含量在0.15~0.57ng/L范围内,而人体尿液中雌二醇含量在0·75~135μg/L范围内。  相似文献   

5.
建立了蓖麻毒素的3种酶联免疫定量分析法,即光吸收、荧光和化学发光免疫分析法,并系统优化了各项实验条件。结果显示:对于化学发光检测法,当酶标抗体HRP-4C13稀释倍数为1∶8000时可获得最佳的信噪比,且酶与底物反应3 min后信号趋于稳定。随后在各自优化的条件下将3种方法用于毒素的检测。比较结果表明:化学发光酶联免疫分析法除具有线性范围宽(0.02~5.5μg/L,r2=0.999)、灵敏度高(检出限为5 ng/L)的特点外,还具有简单、快速、体系稳定性好等优点。将本方法用于不同实际样品基质,如饮用水、碳酸饮料、奶粉、咖啡和血清中添加的蓖麻毒素的检测,其检出限为0.005~0.08μg/L,回收率为89.6%~108.8%,适于污染及中毒样品中痕量蓖麻毒素的定量分析。  相似文献   

6.
建立了一种灵敏的定量检测人碱性成纤维细胞生长因子(hbFGF)脂质体中hbFGF含量的酶联免疫(ELISA)方法,应用于测定hbFGF脂质体的包封率。通过用hbFGF标准品溶液包被酶标板,加入特异性的鼠抗人hbFGF抗体(一抗)和山羊抗鼠IgG(二抗),采用多因素棋盘滴定方法确定最佳实验条件为:一抗浓度200μg/L,二抗浓度50μg/L。本方法的检出限为0.86μg/L,工作浓度范围1.0~10μg/L,批内RSD为3.95%~4.27%,批间RSD为4.19%~7.21%,不同操作者批内和批间的RSD分别为6.21%和9.12%,可重复性良好。本方法测定hbFGF脂质体超滤液和氯仿提取液中hbFGF含量分别为(3.325±0.193)μg/L和(18.454±1.063)μg/L,组内RSD分别为4.69%和4.52%。hbFGF脂质体包封率为82.06%。本方法特异性高、重复性好、快速、简便,可用于hbFGF脂质体包封率及其它含hbFGF制剂的含量测定。  相似文献   

7.
以磁性酶免疫测量分析为模型,建立了微板式磁化学发光酶免疫分析法.利用3-(2′-螺旋金刚烷)-4-甲氧基-4-(3″-磷酰氧基)苯-1,2-二氧杂环丁烷(AMPPD)-碱性磷酸酶(ALP)化学发光体系对人绒毛膜促性腺激素(HCG)进行测定,测量的灵敏度较分光光度法提高了13倍.测定的线性范围为0.15~500 mIU/mL;批内变异系数(C.V.%)和批间变异系数(C.V.%) 均在18%之内;回收率在80%~116%之间;利用本法对血清样品进行了测定,并与其它化学发光免疫分析方法进行了比较,其相关系数为0.965.首次将酶免疫分析方法用于唾液中人绒毛膜促性腺激素(HCG)的测定,为建立非侵入式样品测定方法打下了基础.  相似文献   

8.
化学发光免疫分析法检测血清中前列腺特异抗原复合物   总被引:1,自引:0,他引:1  
采用杂交瘤技术构建抗前列腺特异抗原-α1-抗胰凝乳蛋白酶( Prostate specific antigen-α1-antichymot-rypsin complex,PSA-ACT)单克隆抗体(单抗)细胞株,共获得8株稳定分泌PSA-ACT单抗的细胞株,纯化后进行免疫特性鉴定并做单抗配对。选择其中一对单抗,采用辣根过氧化物酶( HRP)催化鲁米诺( lumino)-H2 O2化学发光体系,建立了测定人血清中PSA-ACT浓度的化学发光酶免疫分析方法( CLEIA)。对包被缓冲液、包被抗体浓度、酶标抗体稀释度、温育时间、发光反应时间和免疫反应步骤等实验参数进行了优化。在最佳实验条件下, PSA-ACT浓度在5~40 ng/mL范围内线性关系良好(R2=0.9943),检出限为0.53 ng/mL,批内相对标准偏差(RSD)为4.6%~6.6%,批间RSD为5.7%~8.0%,回收率为95.4%~104.2%,与游离前列腺特异性抗原F-PSA的交叉反应率为0.6%。本方法简单、稳定、灵敏、快速,为开发检测PSA-ACT的CLEIA试剂盒奠定了基础。  相似文献   

9.
刘海生  刘伟  章竹君 《分析化学》2005,33(6):811-813
设计了一种将微注样阀和发光试剂均集成化的微流动注射化学发光芯片。利用luminolK3Fe(CN)6H2O2化学发光体系,研究了这种芯片的分析特性。该芯片测定H2O2的线性范围为2×10-5~8×10-9mol/L;检出限为3.6×10-9mol/L;相对标准偏差RSD=4.4%(c=1×10-6mol/L,n=11)。与常规的流动注射化学发光分析法相比,该芯片具有简单、快速、灵敏度高、耗样量少等特点,并结合酶促反应成功地用于人体血清中葡萄糖的测定。  相似文献   

10.
待测尿液中的雌三醇、辣根过氧化物酶标记的雌三醇与异硫氰酸荧光素(FITC)标记的兔抗雌三醇抗体在均相体系中发生竞争性免疫反应,再加入用羊抗FITC抗体包被的磁微粒,反应生成物结合在磁微粒上,在磁场中经分离、洗涤后加发光底物,检测发光强度(RLU),测定尿液中雌三醇的含量。通过对检测条件的优化,建立了磁性微粒子化学发光免疫分析法测定人尿液中雌三醇的方法,并对正常男性、女性和孕妇的尿液中雌三醇含量进行了测定。结果表明,本方法的线性范围为1~100μg/L,检出限为0.25μg/L,具有很高的灵敏度;批内相对标准偏差<14%;批间相对标准偏差<7%,具有良好的稳定性和重现性。  相似文献   

11.
用溶胶-凝胶法以磷钼酸(MPA)的镍盐溶液水解钛酸四丁酯制备了NiPMo/TiO2催化剂.使用ICP、 XRD、 TG-DTA、 IR、 TPD-MS和微反应技术研究了催化剂的化学组成、热稳定性、化学吸附性质和催化反应性能.杂多钼酸盐与TiO2通过O2-在TiO2表面发生了键合.在623 K下,杂多阴离子仍保持原有的Keggin结构.CO2在Lewis酸位Ni(Ⅱ)和Lewis碱位Ni-O-Mo的桥氧协同作用下生成CO2卧式吸附态Ni(Ⅱ)←O-(CO)←(O--Ni).丙烯有多种吸附态在催化剂上吸附.在563 K、 1 MPa和空速1500 h-1的反应条件下,丙烯的摩尔转化率为3.2%,产物MAA选择性为95%.  相似文献   

12.
Different approaches for the synthesis of 1-benzyloxypyrazin-2(1H)-one derivatives from simple amino acids have been investigated. A library of 33 precursors for the preparation of N-hydroxy pyrazinones was obtained in moderate to good yields.  相似文献   

13.
In the context of the preparation of camptothecin and luotonin A analogs, the synthesis of some key keto-precursors and their use in Friedländer condensation are described. This paper also focuses on the stability of these keto intermediates and emphasizes the major differences between indolizinones and pyrroloquinazolinones series. Noteworthy is also the report of some original structures isolated as by-products of some experiments.  相似文献   

14.
The Langevin paramagnetic theory can’t describe the relation between magnetization of ferrofluids and applied magnetic field. The structuralization of ferrofluids, which is considered the main influence factor of the magnetization, is regarded. The part of magnetization works is deposited when the structure is forming. This action influences the magnetization of ferrofluids directly or indirectly. On the base of the “compressing” model, the Langevin function that usually describes the magnetization of ferrofluid is modified, and a well-fitted curve is obtained. An equation of the relation between the equivalent volume fraction after being “compressed” and the intensity of magnetic field is discovered, which approximately describes the process of magnetization. The relation between the approximate initial susceptibility and the volume fraction can be obtained from modified formula.  相似文献   

15.
The highly regioselective Buchwald–Hartwig amination at C-2 of the cheap and readily accessible reagent, 2,4-dichloropyridine with a range of anilines and heterocyclic amines is described. This new methodology is robust and provides a facile access to 4-chloro-N-phenylpyridin-2-amines on 0.25 mol scale. These intermediates undergo a further Buchwald–Hartwig amination at higher temperature to enable rapid exploration of the chemical space at C-4 and to provide a library of 2,4-bisaminopyridines.  相似文献   

16.
KMnO4-mediated oxidative CN bond cleavage of tertiary amines producing secondary amine was introduced, which was trapped by electrophiles (acyl chloride and sulfonyl chloride) to form amides and sulfonamides. The reaction could take place at mild condition, tolerating a wide range of function groups and affording products in moderate to excellent yields.  相似文献   

17.
The review contains a concise historical account and information on the most significant researches undertaken by the staff at the A. E. Favorsky Irkutsk Institute of Chemistry, Siberian Branch of the Russian Academy of Sciences on the Chemistry of Heterocyclic Compounds. Dedicated to Academician of the Russian Academy of Sciences B. A. Trofimov on his 70th jubilee. Translated from Khimiya Geterotsiklicheskikh Soedinenii, No. 10, pp. 1443–1502, October, 2008.  相似文献   

18.
A general synthesis of previously unknown semicarbazone-based α-amidoalkylating reagents, 4-(tosylmethyl)semicarbazones, has been developed. The synthesis involved three-component condensation of semicarbazones of aliphatic or aromatic aldehydes with the same or other aldehydes and p-toluenesulfinic acid. The scope and limitations of this reaction were investigated. The compounds obtained were demonstrated to be an efficient α-(4-semicarbazono)alkylating agents. They were reacted with H- (sodium borohydride), O- (sodium methylate), S- (sodium phenylthiolate), N- (pyrrolidine, sodium succinimide), P- (trialkyl phosphites), and C-nucleophiles (sodium diethyl malonate) to give the corresponding products of the tosyl group substitution, 4-substituted semicarbazones, including analogues of nitrofurazone. Among the prepared compounds tested in vitro for antibacterial and antifungal activity, three nitrofuryl-containing semicarbazones exhibited high biological activities with minimum inhibitory concentration (MIC) values of 8–32 μg/mL.  相似文献   

19.
Zhanhui Yang  Shiyi Yang  Jiaxi Xu 《Tetrahedron》2017,73(23):3240-3248
Regiospecific and direct imidation of the methyl C(sp3)–H bond of thioanisoles is realized under mild and metal-free conditions with N-fluorobis(benzenesulfonyl)imide as an oxidant and nitrogen source. Proposed mechanism suggests that thionium ion intermediates and a Pummerer-type reaction are involved. The imidation has advantages such as high step-economy, excellent functionality tolerance, and regiospecificity, giving structurally diverse imidation products.  相似文献   

20.
A small library of new chiral bidentate hydroxyalkyl-imidazolium salts 1 is conveniently synthesized on multi-gram scale from inexpensive and commercially available chiral pool amino acids. The corresponding carbenes, generated by deprotonation of imidazolium salts 1, in combination with palladium(II) chloride were tested in the Mizoroki–Heck coupling reaction. The most significant results in terms of yields and reactivities were achieved with low catalyst loading. The catalytic activities of these imidazolium salts were also investigated in the asymmetric addition of diethylzinc to benzaldehyde. The use of MgO nanoparticles as an additive in conjunction with these ligands played a crucial role in increasing the efficiency of these reactions.  相似文献   

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