双抗体夹心ELISA法检测海产蟹血卵涡鞭虫方法的初步建立

以针对血卵涡鞭虫的特异性单克隆抗体为捕获抗体,纯化的抗血卵涡鞭虫的兔抗血清为检测抗体,初步建立了检测血卵涡鞭虫的双抗体夹心ELISA方法.方阵滴定确定最佳反应条件:包被的单克隆抗体浓度为10μg/mL,多抗兔血清工作浓度为14.3 μg/mL,待检样品稀释度为1:800;HRP-羊抗鼠IgG按1:20 000稀释.用建立的ELISA方法对86份已知背景样品进行检测,阳性检出为97.6%,血卵涡鞭虫抗原检测的灵敏度为100 pg/mL;与蟹血淋巴细胞、假丝酵母菌、溶藻弧菌、副溶血弧菌等样本均无交叉反应.结果表明,建立的双抗体夹心ELISA方法灵敏度高、特异性强,可用于诊断梭子蟹乳化病血卵涡鞭虫病的检测.     

改进双抗夹心ELISA用于筛选单抗杂交瘤细胞株

本文利用改进双抗夹心ELISA筛选一种可测定血清样品中与血浆蛋白结合的蛋白质药物的单抗。用兔抗K102多抗经亲和层析纯化并包被后,加入用10%猴血浆PBS还原后的抗原K102,再加入杂交瘤细胞株上清液和HRP-羊抗鼠IgG、底物反应液筛选阳性单抗杂交瘤细胞株。结果用改进后的双抗夹心ELISA筛成功地选出6株能稳定分泌针对K102的单抗杂交瘤细胞株,分别命名为1C9、2D7、3E2、5F4、5F6、6B9;细胞株产生的单抗用ELISA法能检测出血清样品中的K102的浓度。提示改进的双抗夹心ELISA可用于针对与血浆蛋白结合的蛋白质药物的单抗杂交瘤细胞株的筛选。     

BK病毒样颗粒制备及血清学检测方法的建立

将BK病毒 ( BK polyomavirus, BKV )的结构蛋白VP1 通过重组杆状病毒在昆虫细胞中表达, 并自发装配成BKV病毒样颗粒(BK virus like particles, BK VLPs). 以BK VLPs作为包被抗原建立了间接酶联免疫吸附试验(enzyme linked immunosorbent assay, ELISA)的血清学检测方法: 抗原最佳包被浓度为3.1 μg/mL, 血清最佳稀释度为1∶200, 酶标抗体最佳工作稀释度为1∶40000.并以该方法在国内对540名健康成年人进行BKV抗体阳性率情况调查, 结果显示我国健康成年人BKV抗体阳性率为79.94%.     

脊髓灰质炎病毒Ⅰ型抗体胶体金检测试纸条的研制

目的:为研制快速检测脊髓灰质炎病毒Ⅰ型抗体的胶体金试纸条.方法:以辛酸-硫酸铵沉淀法纯化脊髓灰质炎病毒Ⅰ型单克隆抗体,用柠檬酸三钠还原法制备胶体金溶液,胶体金标记单抗,再结合脊灰病毒抗原形成复合物,固定于金标垫上,将鼠抗人IgG和羊抗鼠IgG分别包被于试纸条的T (检测线)处和C (质控线)处,应用该试纸条进行性能检测.结果:以已知的脊灰病毒Ⅰ型阳性血清作为检测样品,试纸条检测的灵敏度为1:80;特异性试验证明该试纸条与甲型肝炎病毒、乙型肝炎病毒、麻疹病毒抗体阳性血清没有交叉反应;检测结果与脊灰病毒Ⅰ型疫苗抗体检测试剂盒(ELISA)试验的符合率为86.7%.结论:该试纸条检测脊灰病毒Ⅰ型的抗体水平具有较好的特异性与灵敏性,适用于大规模临床血清抗体水平检测,具有良好的应用前景.     

鳗鲡源创伤弧菌的间接ELISA快速检测

创伤弧菌(Vibrio vulnificus)血清2型对鳗鲡有专一的感染性,是鳗鲡出血性败血症、烂鳃病和烂尾病的重要病原菌。采用间接ELISA法对制备的兔抗创伤弧菌多克隆抗血清进行效价测定,结果表明其效价高达1∶512 000。从发病鳗鲡中分离出创伤弧菌和37株其他致病菌,将38株病原菌以较低浓度(1×106cfu/m L)包被后通过兔抗创伤弧菌血清进行交叉反应水平检测,结果表明,兔抗创伤弧菌血清稀释到1∶256 000或1∶128 000时,其与37株其他病原菌均未产生明显的ELISA交叉反应。该血清分别经1∶64 000和1∶32 000稀释后,采用ELISA方法检测了经创伤弧菌、鳗鲡爱德华氏菌(Edwardsiella anguillarum)和嗜水气单胞菌(Aeromonas hydrophila)攻毒感染死亡的美洲鳗鲡(Auguilla rostrata)脏器组织悬液(肝脏、肾脏和鳃),结果表明,兔抗创伤弧菌血清经1∶32 000稀释后可特异性检出鳗鲡鳃和肾脏组织中的创伤弧菌。     

轮状病毒间接免疫荧光的检测方法

以兔抗SA11多克隆抗体为一抗、 Alexa 488标记的山羊抗兔抗体为二抗, 通过对反应条件的优化建立轮状病毒（RV）检测的间接免疫荧光法（IFA）. 实验结果表明, IFA最佳工作条件为: MA104细胞密度为每孔2×10⁴个, 胰酶活化轮状病毒质量浓度为1 μg/mL, 培养时间为18 h, 一抗最适稀释度为1∶800, 二抗最适稀释度为1∶500.     

KGF-2单克隆抗体间接ELISA筛选方法的建立

目的建立KGF-2单克隆抗体间接ELISA的筛选方法 ,制备KGF-2单克隆抗体.方法以KGF-2为抗原免疫BALB/c小鼠,获得抗血清.通过棋盘滴定方法确定间接ELISA的抗原包被浓度及一抗、二抗最佳工作浓度;利用建立特异性好的间接ELISA方法筛选杂交瘤细胞株.结果通过棋盘滴定最终确定了500μg/L的抗原包被浓度、1∶1 000的一抗血清稀释倍数及1∶2 000的二抗血清稀释倍数;筛选到两株可稳定分泌抗体、效价较高的杂交瘤细胞株.结论该研究建立了灵敏度高、特异性好的间接ELISA筛选体系,可用于KGF-2单克隆抗体的制备及检测.     

鸡传染性支气管炎病毒抗体胶体金免疫层析试纸条的研制

[目的]建立一种快速、灵敏的胶体金免疫层析法(Colloidal-gold immunochromatography assay),用以检测禽类早期感染鸡传染性支气管炎病毒(Infectious bronchitis virus,IBV)和监测IBV疫苗接种免疫水平.[方法]将重组IBV核衣壳(Nucleocapsid,N)蛋白包被在在试纸条检测线,羊抗兔IgG包被在试纸条控制线,用胶体金-兔抗鸡IgG偶联物作为示踪剂,检测血清中IBV抗体.[结果]制备所得的胶体金免疫层析试纸条与IBV抗体阳性血清反应呈阳性,与鸡新城疫病毒抗体阳性血清、鸡传染性法氏囊病毒抗体阳性血清、鸡毒霉形体抗体阳性血清和SPF标准鸡血清未发生交叉反应;当IBV抗体阳性血清以1:35比例稀释后,依然可以用该试纸条检出;该试纸条在60 d的保存期内重复性良好;以从不同养殖场采集到的鸡血清样本作为测试对象,比较该试纸条和进口 ELISA试剂盒的检测结果,两者符合率为97.44％.[结论]成功制备了检测耗时短、检测限低、特异性强、稳定性好的IBV抗体检测胶体金免疫层析试纸条,为鸡传染性支气管炎的早期诊断以及免疫监测提供了简便高效的技术方法.     

大肠埃希菌ELISA检测条件的优化

目的对ELISA检测大肠埃希菌方法条件进行优化研究.方法以大肠埃希菌为对象,通过Box-Behnken设计星点响应面实验进行优化实验.结果优化后ELISA检测条件:抗原包被浓度为6.25μg/m L,抗原包被最佳时间为7 h,抗体稀释度最佳为1∶625 0,大肠埃希菌ELISA检测OD值为2.12.结论经实验得证,通过星点响应面实验方法进行大肠埃希菌ELISA检测条件优化,结果合理有效,优化效果良好.     

应用酶联免疫吸附试验检测山羊日本血吸虫病抗体的研究

本研究首先制备了日本血吸虫水溶性止卵抗原和兔抗山羊酶标复合物。经测试确定了:抗原的最适包被浓度(5ug/ml);酶标复合物的最佳工作浓度(1:2500)以及血清不同稀释度下的阴—阳临界值,从而成功地建立了山羊日本血吸虫病抗体的酶联免疫检测法。以1:80稀释作参考,对25份标准阳性血.清和19份标准阴性血清的考核结果证实:在最适条件下,该法特异性,敏感性良好,具有临床和科研应用价值。随后,应用酶联免疫吸附试验对7只实验山羊血吸虫病抗体消长作了长期、细致地观察,结果表明:特异性IgG最早于尾蚴感染后第40天出现;吡喹酮治疗后,7个月消失。     

Markedness and its Interpretation in Language Use

Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.     

Features of Women's language

Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women＇s language from these perspectives： pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women＇s language, more attention should be paid to language use in social context. What＇s more, the linguistic phenomena in a speaking community can be understood more thoroughly.     

低渗透非均质砂岩油藏启动压力梯度研究

理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。     

On Tragic Consciousness In The Works By Ernest Hemingway

As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.     

Achieving Unusual States of Matter Under High Pressure

正The periodicity of the elements and the non-reactivity of the inner-shell electrons are two related principles of chemistry,rooted in the atomic shell structure.Within compounds,Group I elements,for example,invariably assume the+1 oxidation state,and their chemical properties differ completely from those of the p-block elements.These general rules govern our understanding of chemical structures and reactions.Using first principles calcula-     

Exchange-Correlation and Electronic Excitation Energies from Pairing Matrix Fluctuations

We have developed an adiabatic connection to formulate the ground-state exchange-correlation energy in terms of pairing matrix linear fluctuations.This formulation of the exchange-correlation energy opens a new channel for density functional approximations based on the many-body perturbation theory.We illustrate the potential of such approaches with an approximation based on the particle-particle Random Phase Approximation(pp-RPA).This re-     

Density Functional Theory for the Response of Periodic Systems to Electric Fields Based on the Vector Potential Approach

正The electronic and nuclear(structural/vibrational)response of 1D-3D nanoscale systems to electric fields gives rise to a host of optical,mechanical,spectral,etc.properties that are of high theoretical and applied interest.Due to the computational difficulty of treating such large systems it is convenient to model them as infinite and periodic(at least,in first approximation).The fundamental theoretical/computational problem in doing so is that     

On the Thermodynamic Limit for Responses to Static Electromagnetic Fields

For molecular systems,the quantum-mechanical treatment of their responses to static electromagnetic fields usually employs a scalar-potential treatment of the electric field and a vector-potential treatment of the magnetic field.Although the potential for each field separately is associated with the choice of an(unphysical)origin,the precise choice of the origin for the electrostatic field has little consequences for the results.This is different for the     

An affordable accurate evaluation of the binding conformations predicted by molecular docking studies in bio-system

Franck-Condon factors bridge the gap between theoretical modeling and experimental observations for molecular electronic spectroscopy and electron transfer.Under the displaced harmonic oscillator approximation,multidimensional Franck-Condon factors are decomposed into a product of many one-dimensional(1D)Franck-Condon(FC)factors,and each 1D-FC factor is associated with one Huang-Rhys factor that determines the leading contribution of     

Call for Papers

<正>"The Journal of Shanghai Normal University:Mathematics"is published by Shanghai Normal University as regular issues of The Journal of Shanghai Normal University each year from 2014 in English.The editors-in-chief of the issues are professors Yuhao Cong and Maoan Han.The Journal of Shanghai Normal University was started in 1958 with