胶体金免疫层析试纸条在食品安全检测中的研究进展

为了提高免疫检测的速度,简化操作步骤,建立了胶体金免疫层析技术.它是利用纤维膜的毛细作用,使待测物和金标物(抗原/抗体)沿膜表面向前运动,与检测区的捕获物特异结合而聚集显色.它既有免疫学方法的特异性,又有简便快速,无需仪器的优点.本文主要介绍了胶体金免疫层析技术的在食品安全检测中的应用,并对胶体金免疫层析试纸条技术的发...     

呋喃唑酮代谢物单克隆抗体及其新型免疫层析检测试纸条的制备

目的制备呋喃唑酮代谢物AOZ单克隆抗体及其胶体金与荧光淬灭免疫层析试纸条。方法将AOZ衍生成CPAOZ后,与载体蛋白BSA偶联,制备完全抗原CPAOZ-BSA,经皮下多点注射免疫BALB/c小鼠,共5次,取小鼠脾细胞与SP2/0细胞融合,获得稳定分泌特异性抗体的杂交瘤细胞株。利用获得的抗CPAOZ单克隆抗体制备胶体金及荧光淬灭免疫层析试纸条,并对试纸条的灵敏度及特异性进行验证。结果制备的完全抗原经紫外扫描鉴定偶联成功。共获得3株能稳定分泌抗CPAOZ单克隆抗体的杂交瘤细胞株,其中效价和特异性最好的1株2H11针对CPAOZ的50%抑制质量浓度(IC50)为0.88 ppb。以该抗体制备的胶体金及荧光淬灭免疫层析试纸条的最低检测限分别为7.5和0.062 5 ppb;两种试纸条与其他3种硝基呋喃类代谢物CPAMOZ、CPAHD、CPSEM均不存在交叉反应。结论成功制备了抗CPAOZ单抗及其胶体金免疫层析和荧光淬灭免疫层析试纸条,两种试纸条均具有较高的灵敏度和较强的特异性,其中荧光淬灭免疫层析试纸条的灵敏度较胶体金免疫层析试纸条高约100倍,具有良好的应用前景。     

三聚氰胺快速检测试纸条评价

胶体金试纸条因其方法简便、快速,已经广泛应用于日常检测和实验研究中.为了解国产三聚氰胺试纸条的有效性和可靠性,选择了一种市售试纸条,从灵敏度、特异性、准确度、耐变性、批间变异等多个角度对其进行了技术评价.结果发现用胶体金法测定牛奶中的三聚氰胺,检测限为0.05 mg/L,特异性、耐变性较好,批件差异较小,不同基质对检测结果无影响.此试纸条满足食品中有毒有害物质快速检测的要求,可作商品化快速检测试纸条使用.     

玉米细菌性枯萎病菌纳米磁标记免疫层析检测试剂与试纸条

分别以粒径为50和100 nm的羧基化纳米磁珠,通过碳二亚胺(EDC)/N-羟基琥珀酰亚胺(NHS)交联法使磁珠与抗体偶联,制备出可特异性识别玉米细菌性枯萎病菌的两种纳米免疫磁珠试剂。基于双抗体夹心法,经实验条件优化,选用100 nm的羧基化纳米磁珠为磁标记探针,组装了玉米细菌性枯萎病菌磁标记免疫层析检测试纸条。该方法用于玉米种子中玉米细菌性枯萎病菌的检测,检出限为1.0×106CFU/m L,15 min内可通过肉眼判读结果,也可用超顺磁共振检测仪实现量化检测,具有简便、快速、现场化检测的优势。     

胶体金试纸条法快速检测纺织品中总铅和总镉含量

建立了一种基于铅、镉单克隆抗体的胶体金试纸条方法,用于快速检测纺织品中总铅、总镉含量,通过加标实验及对实际试样进行测试,探讨了试纸条的灵敏度、准确性和稳定性.结果表明:制备胶体金试纸条时,铅、镉单克隆抗体较佳用量分别为50,45μL;试纸条对纺织品中总铅及总镉含量的检出限分别达5.0 mg/kg和0.625 mg/kg...     

ELISA法与胶体金试纸条检测HBsAg差异分析

目的探究胶体金试纸条与ELISA法检测HBsAg临床应用价值。方法分别用胶体金试纸条与ELISA法检测360份标本中的HBsAg,并对阳性率进行比较,然后将强阳性的10份标本作倍比稀释,观察2种检测方法灵敏度的差异。结果2种方法的阳性率之间无显著差异,ELISA法灵敏度较胶体金试纸条灵敏度高。结论2种方法皆是检测HBsAg的良好方法,胶体金试纸条法操作简便、快速适用于急诊及健康人群的筛查,ELISA法适用于常规检测。     

白斑综合征病毒胶体金免疫层析试剂板的研制

目的研制检测对虾中白斑综合征病毒(white spot syndrome virum,WSSV)的胶体金免疫层析试剂板。方法分别制备抗WSSV单克隆抗体和多克隆抗体,并用胶体金标记单克隆抗体,将胶体金标记的抗WSSV单克隆抗体包被在金标垫上,抗WSSV多克隆抗体和羊抗鼠Ig G分别包被在硝酸纤维素膜的检测线和质控线上,组装成试剂板,通过双抗体夹心法检测样品中WSSV含量。确定试剂板检测WSSV的最低检出限,验证其特异性,并检测其与ELISA结果的符合率及稳定性。结果该试剂板对WSSV的最低检出限为1.0 mg/kg,特异性强,与ELISA实验结果相符,在室温条件下(20~25℃)可保存12个月。整个检测所需时间为8~10 min。结论该试剂板能快速有效地筛查WSSV,适合现场快速检测。     

犬细粒棘球绦虫感染胶体金检测试纸条的制备

目的制备犬细粒棘球绦虫感染胶体金检测试纸条。方法采用柠檬酸三钠还原法制备胶体金颗粒,用于标记抗细粒棘球绦虫Ediag A864单克隆抗体2D12制作金标垫,兔抗细粒棘球绦虫Ediag A864多克隆抗体标记检测线(T线),羊抗鼠Ig标记质控线(C线),制备胶体金试纸条,并进行特异性、敏感性、重复性及稳定性试验。用优化的胶体金检测试纸条对96份犬粪样品(36份参照阳性样,60份临床样品)进行检测。结果制备的犬细粒棘球绦虫感染胶体金检测试纸条与犬蛔虫阳性粪及犬贾第虫阳性粪样品均无交叉反应;粪便稀释度为1∶4时仍可检出正确结果;3次检测参照阳性样本均为阳性;于不同温度(室温、4及37℃)下保存不同时间(1、2、3、4个月)的试纸条均可检出正确结果。96份犬粪便样品中,36份参照阳性样品检出率为97.2%(35/36),60份临床样品均为阴性。结论制备的试纸条具有良好的特异性及稳定性,可应用于临床样品检测及大规模流行病学调查。     

布病胶体金免疫层析检测方法的建立

目的建立诊断布病的胶体金免疫层析方法。方法以B.abortus 544A全菌免疫BALB/c小鼠,以杂交瘤细胞技术制备单抗,纯化后以胶体金标记,选择最佳反应模式组装试纸条,并对其进行鉴定。结果已筛选4B8、1G9、1C9和1E24株稳定分泌单抗的细胞株,以其单抗互相配对组合的试纸条均能检出B.abortus 544A菌。其中以1G9为包被抗体、4B8为金标抗体的试纸检测效果最佳,与以B.abortus 544A菌多抗为包被抗体、4B8单抗为金标抗体的试纸对比,检测结果一致,且不与其他菌发生交叉反应。对PCR检测为阳性的70份牛血清样品,两种试纸条检测结果的符合率均为100%。结论所建立的诊断布病的胶体金免疫层析法快速、简便、准确,有望用于布病的诊断。     

胶体金层析免疫法对盐酸克伦特罗的检测试验

盐酸克伦特罗俗称"瘦肉精",若大量添加到猪饲料中,将在猪肉及内脏中残留,给食品安全带来了隐患。传统的检测技术繁琐,成本高,不利于推广普及。本实验利用胶体金层析法检测盐酸克伦特罗,结果表明:最低检测限度为5 ppb,具有简便、便捷、快速、灵敏度强的特点。     

胶体金免疫层析法检测小分子物质

在过去的6年里,胶体金及免疫层析试剂越来越多的被用于快速检测上。这一简单免疫层析试剂给很多不同分析物提供了简便而又快速的方法,它的出现给很多工业带来了冲击。最近,胶体金免疫层析法检测小分子物质的报道日益增多,特别是对兽药和农药的检测。文章主要讲述了用于检测小分子物质的胶体金检测试纸的制备原理、检测原理、应用和进展。     

一种新型氰戊菊酯农药人工抗原的合成鉴定与免疫效果的探索

文献报道的氰戊菊酯半抗原结构有多种,分别从氰戊菊酯二苯醚和α-氰基部位引出连接臂合成人工抗原。为了探寻抗体与目标分子互作取向规律,以氰戊菊酯为载体,基于半抗原不同连接臂位点,同时也考虑到二苯醚和α-氰基结构为多种菊酯农药所共有,因此从其特征结构氰戊菊酸苯环结构上引出连接臂位点设计了一种新半抗原,采用α-异丙基苯乙腈等为主要原料,分别经过6和7步反应合成了两种连接臂不同的新结构的半抗原,并采用质谱和核磁共振手段对该新化合物进行结构表征;然后通过重氮化和活泼酯法将两种半抗原共价偶联到牛血清白蛋白(BSA)和卵清蛋白(OVA),经过紫外光谱扫描、电泳进行鉴定,证明偶联成功,免疫小鼠结果 IC50值为δ2.4,与相关拟除虫菊酯农药有一定交叉反应,为抗原抗体识别位点和机制的探索提供了新的数据支撑。     

Development of a Rapid Fluorescent Diagnostic System for Early Detection of the Highly Pathogenic Avian Influenza H5 Clade 2.3.4.4 Viruses in Chicken Stool

Rapid diagnosis is essential for the control and prevention of H5 highly pathogenic avian influenza viruses (HPAIVs). However, highly sensitive and rapid diagnostic systems have shown limited performance due to specific antibody scarcity. In this study, two novel specific monoclonal antibodies (mAbs) for clade 2.3.4.4 H5Nx viruses were developed by using an immunogen from a reversed genetic influenza virus (RGV). These mAbs were combined with fluorescence europium nanoparticles and an optimized lysis buffer, which were further used for developing a fluorescent immunochromatographic rapid strip test (FICT) for early detection of H5Nx influenza viruses on chicken stool samples. The result indicates that the limit of detection (LoD) of the developed FICT was 40 HAU/mL for detection of HPAIV H5 clade 2.3.4.4b in spiked chicken stool samples, which corresponded to 4.78 × 10⁴ RNA copies as obtained from real-time polymerase chain reaction (RT-PCR). An experimental challenge of chicken with H5N6 HPAIV is lethal for chicken three days post-infection (DPI). Interestingly, our FICT could detect H5N6 in stool samples at 2 DPI earlier, with 100% relative sensitivity in comparison with RT-PCR, and it showed 50% higher sensitivity than the traditional colloidal gold-based rapid diagnostic test using the same mAbs pair. In conclusion, our rapid diagnostic method can be utilized for the early detection of H5Nx 2.3.4.4 HPAIVs in avian fecal samples from poultry farms or for influenza surveillance in wild migratory birds.     

20%氰戊菊酯·马拉硫磷乳油的气相色谱程序升温法同时分析

建立气相色谱程序升温法同时分析氰戊菊酯和马拉硫磷含量的方法。色谱柱为Rtx-1毛细管色谱柱,癸二酸二正丁酯为内标物,丙酮为溶剂。实验表明,氰戊菊酯和马拉硫磷的线性相关系数分别为0.996 4和0.998 4,变异系数分别为0.308 6和0.240 1,平均回收率分别为99.57%和99.78%。     

镀金添加剂的研究进展

回顾了镀金添加剂的研究概况,介绍了镀金添加剂的分类及应用,提出了镀金添加剂今后的研究重点。     

Electrochemical sensor-based gold nanoparticle/poly(aniline-co-o-toluidine)/graphene oxide nanocomposite modified electrode for hexavalent chromium detection: a real test sample

This paper presents the fabrication of poly(aniline-co-o-toluidine)/graphene oxide nanocomposite with a general abbreviation [PANI-co-PoT/GO_a–e] by well-known in-situ oxidative polymerization method with ultrasonic assistance. These materials were based on variable loading of GO when prepared. The chemical structures of the composite materials were confirmed by characterization technique. The FE-SEM and TEM micrographs were used to investigate the morphological features. Furthermore, FT-IR, XRD, TGA, and electrical conductivity measurements were utilized to estimate its complete performance. All nanocomposites showed CDT_max values in the range of 287.25–463.51 ºC which is significantly higher than that observed for pure copolymer (204.79 ºC). The main focus of this paper is to study the electroselective application using gold nanoparticle as a coating. A steady electroactive modified electrode [AuNPs/PANI-co-PoT/GO] was effectively prepared on a gold electrode (Au) surface using an electroadsorption process for the determination of Cr(VI). The electrochemical attitude of the modified sensor toward the reduction of Cr(VI) was studied by a square wave voltammetry (SWV) and a cyclic voltammetry (CV) technique. The AuNPs/PANI-co-PoT/GO modified electrode displayed a perfect electrochemical activity toward the reduction of Cr(VI). Using an SWV method, the modified electrode gave a linear response to Cr(VI) through the concentration range 5–500 µM with a limit of detection 0.0215 µM. The suggested sensor displayed good stability, sensitivity and selectivity and has exhibited potential for the detection of Cr(VI) in real samples.     

胶体金技术测定黄瓜中甲霜灵残留

比较了胶体金试纸法与气相色谱法在检测黄瓜甲霜灵残留中的应用。结果表明,胶体金试纸检测为阴性的样品,经气相色谱检测结果为未检出,阳性样品经气相色谱检测结果与胶体金试纸相一致。与气相色谱法比较,胶体金试纸法检测甲霜灵具有操作简便、直观、快速、省时的特点,其特异性、较高的敏感性,可作为黄瓜中甲霜灵残留现场检测的手段应用。     

用于汽车密封条的HPVC热塑性弹性体的研制

考察了3个不同厂家HPVC的基本性能,优选出适合制备弹性体的树脂,再通过加入增塑剂、与橡胶共混以及交联改性开发出了可用于汽车密封条的HPVC热塑性弹性体。     

微型继电器接触簧片镀金工艺研究

基于传统镀金工艺存在的不足,结合超声波电镀机理,提出了一种新的适合微型继电器接触簧片的镀金工艺.该工艺结合了滚镀与挂镀的特点,采用自制简易振动篮。通过实验确定了时间、电流和镀层厚度之间的关系,并发现采用该工艺得到的镀层能满足产品使用要求,指出了该工艺维护的要点及不足。     

Preparation and characterization of polyurethane–gold nanocomposites prepared using encapsulated gold nanoparticles

Gold nanoparticles (GNPs) have been widely studied due to their unique properties. Although many research groups have developed the synthesis of GNPs using various polymers as stabilizing or reducing agents, the effects of GNPs on the structures and properties of polymer matrices have been less reported. We propose a new design for the preparation of polyurethane–gold (PU–Au) nanocomposites. 11‐Mercapto‐1‐undecanol‐coated GNPs acted as the chain extenders and reacted with isocyanates to form covalent bonds between PU and GNPs. PU–Au nanocomposites were successfully synthesized, and the effects of multifunctional GNPs on the structures, morphology and properties of poly(ester urethane) were investigated. Scanning electron microscopy images suggested the GNPs can be dispersed uniformly in the PU matrix. Maltese‐cross of spherical crystals was observed in the PU–Au nanocomposites, and the size of the crystals decreased with an increase in gold content. As the gold content increased, the thermal decomposition temperature and the temperature of the maximum decomposition rate increased. The glass transition temperature, crystal melting temperature and melting enthalpy of the soft segment also increased progressively. The results showed that multifunctional GNPs concentrated hard segments and resulted in an increase of heterogeneous nucleation, phase separation and elasticity. Copyright © 2010 Society of Chemical Industry