简述抗原抗体的体外检测技术

结合教材对抗原抗体检测的描述,从酶联免疫吸附试验、免疫层析技术、免疫印迹技术、化学发光免疫分析以及免疫PCR技术5个方面分析了几种常见的抗原抗体体外检测技术,提出相应的教学建议,拓宽学生的知识视野。     

三种不同包被抗原用于RF的测定与比较

分别用人、兔、鼠IgG作包被抗原,检测244份血清标本(类风湿性关节炎72份,系统性红斑狼疮74份,慢性肾病50份,其它内科疾病33份,健康体检者15份)。类风湿因子RF,发现以AgHIgG作为包被抗原时,方法最敏感,AgRIgG次之,而AgMIgG最差。以AgRIgG为包被抗原(间接法)与以AgHIgG、AgRIgG为包被抗原(双抗原夹心法)所测RF有较好一致性,P分别为0.01,P<0.05,P<0.01。     

皖南地区常见猪病的抗体水平监测与分析

针对皖南地区的部分规模猪场及散养户进行血清学调查,统计了猪瘟、猪蓝耳病、猪口蹄疫、猪圆环病毒病、猪伪狂犬病等常见猪病的抗体阳性率并进行分析,结果显示:(1)四种常见猪病达到国家免疫要求,抗体阳性率≥70%,猪蓝耳病抗体阳性率69.5%,略低于国家免疫要求;(2)皖南各地的猪瘟、猪口蹄疫、猪伪狂犬病的抗体阳性率差异具有统计学意义(P<0.01);(3)不同阶段猪群的猪瘟、猪蓝耳病、猪口蹄疫的抗体阳性率差异具有统计学意义(P<0.01),猪伪狂犬病的抗体阳性率差异具有统计学意义(0.01相似文献   

梅毒螺旋体抗体诊断试剂的临床应用

目的:同时用评价试剂和参比试剂检测436例血清样本,探讨评价试剂与参比试剂检测结果的符合率.方法:评价试剂采用化学发光法,参比试剂采用双抗原夹心酶联免疫吸附试验;并通过卡方检验来判断两试剂盒间有无显著差异.结果:评价试剂和参比试剂阳性符合率为98.25%,阴性符合率为99.47%.两种试剂通过卡方检验比较差异无统计学意义(P>0.05).结论:评价试剂与参比试剂的检测结果具有很好的一致性,评价试剂可以用于梅毒感染的辅助诊断.     

皖南地区规模场猪伪狂犬病血清学分析

针对近年来严重影响规模猪场的传染性疾病猪伪狂犬病,采集了皖南地区31个不同规模种猪场血清样品1000份,使用酶联免疫吸附试验检测法,进行猪伪狂犬病野毒感染抗体血清学调查。结果：31个种猪场中,21个种猪场gE抗体阳性,阳性率67．74％;检测种猪血清1000份,gE抗体阳性率为23％。皖南地区种猪场均有PR感染情况,其中青阳县PR野毒抗体阳性率最高;150头以下的猪场,抗体阳性率较高。分析结果为皖南地区规模猪场制订PR综合防控措施提供了依据。     

植物病毒抗血清的制备及IgG纯化与酶标技术研究

以烟草花叶病毒（TMv）为例研究了制备植物病毒抗血清的一般方法．以家兔和小白鼠为免疫动物，运用饱和硫酸接沉淀法制备出高效价的兔和瓦抗TMV血清，并运用先进的ELISA方法测定了抗血清中抗体IgG的效价；用醋酸纤维凝胶电泳技术分析了抗血清中抗体的lgG纯度，应用免疫双扩散法检测了提纯的IgG的生物活性．同时还运用成二醛一步法制出高结合率的IgG与过氧化氢酶（HRP）标记的抗体，形成了一整套制备抗血清、纯化IgG并对IgG进行酶标的技术，为植物病毒检测提供了快速简便的方法．     

芜湖市规模鸡场禽流感抗体水平的监测与分析

目的:免疫接种是预防禽流感疫情的最主要措施,通过抗体检测判断免疫效果并指导后续免疫措施的改进,为芜湖市禽流感的综合防控提供了基础依据;方法:2013年~2015年期间对芜湖市部分规模鸡场进行血清学调查,采用间接ELISA法(酶联免疫吸附试验)进行检测,对禽流感抗体阳性率进行了分析;结果:(1)2013年至2015年间,芜湖市禽流感免疫情况整体良好,血清抗体阳性率平均达到81.9%,但2014年度春季免疫普查结果低于国家强制免疫要求;(2)抽检的90个养鸡场中,74个抗体阳性率超过国家免疫要求,养鸡场免疫合格率为82.2%;(3)芜湖四区四县中仅弋江区抗体阳性率结果低于国家免疫要求,各区县抽检的血样抗体阳性率差异极显著(P0.01);(4)三种类型养鸡场抗体阳性率存在差异极显著(P0.01),肉鸡场抽检的血样抗体阳性率较低,需针对性加强免疫措施;结论:禽流感抗体水平监测在芜湖市重大动物疫病的防控中起到了积极的预警和指导作用。     

抗雌激素多克隆抗体的制备及效价鉴定

为了快速安全地检测动物性食品中是否含有过高浓度雌激素,本研究将半抗原雌二醇(Estradiol, E₂)合成为雌二醇-3-羧甲基醚(E₂-3-CME),再通过碳二亚胺(EDC)法与牛血清白蛋白(BSA)结合,合成完全抗原(E₂-BSA).将1 mg/只的E₂-BSA免疫雌性家兔,制备出抗雌激素的抗体.通过用酶联免疫吸附(Elisa)实验对产生的抗体进行效价鉴定.实验结果表明,第三次免疫后E₂-BSA产生的抗体光密度值(OD)最高,且与对照组,免疫一次和两次产生的抗体的OD值均具有明显差异,显示通过抗原免疫三次的家兔产生的抗体效价最高,可为后期通过此方法制备抗体和Elisa试剂盒奠定基础.     

张家口市SARS患者、疑似病例、留观及密切接触医护人员血清中IgG、IgM抗体测定

目的 :为临床诊断及排除严重急性呼吸综合征 (SARS)提供依据和对留观及密切接触医护人员有无感染作初步调查。方法 :用酶联免疫吸附测定 (ELISA) 5 3例临床诊断病例、2 9例疑似病例、6 3例留观及 114例密切接触医护人员的血清中SARS冠状病毒IgG、IgM抗体。结果 :SARS临床诊断病例IgG抗体阳性 2 1例 ,IgM抗体阳性 18例 ,其中有 17例IgG、IgM抗体均为阳性 ,总阳性率为 4 1.5 % ;留观人员IgG抗体阳性 1例 ;其他均为阴性。 结论 :ELISA测定血清抗SARS冠状病毒抗体阳性病例可确诊感染病毒 ,对于观察期后 (一般为 2 1d)人员 ,阴性结果可以排除感染。     

放飞思维的个性,激发表达的欲望

<正>一路课改,一路反思。一个个失败的教训,一个个成功的经验,终于让我明白:原来我们需要建设这样的生本课堂——学生自觉学习,积极思考,主动探究,互助交流,有思维的灵动,有智慧的火花,有表达的欲望,能增长智慧,点燃学生成长和生命的课堂。前不久,我在我所执教的八年五班教学臧克家的《有的人》一课。上课伊始,我用这样的问题导入:同学们,学了《藤野先生》一课,你觉得鲁迅是什么样的人?其结果令我大吃     

Development and evaluation of immunoassay for zeranol in bovine urine

A high affinity polyclonal antibody-based enzyme linked immunosorbent assay （ELISA） was developed for the quantification of zeranol in bovine urine. On the basis of urine matrix studies, the optimized dilution factors producing insignificant matrix interference were selected as 1：5 in pretreatment. In the improved ELISA, the linear response range was between 0.02 and 1 μg/ml, and the detection limit was 0.02 μg/ml for the assay. The overall recoveries and the coefficients of variation （CVs） were in the range of 82%-127% and 3.5%-8.8%, respectively. Thirty-six bovine urine samples spiked with zeranol （ranging from 0.2 to 10 μg/ml） were detected by the ELISA and liquid chromatography （LC） method, and good correlations were obtained between the two methods （R^2=0.9643）. We conclude that this improved ELISA is suitable tool for a mass zeranol screening and can be an altemative for the conventional LC method for zeranol in bovine urine.     

Study on the serum levels of soluble intercellular adhesion molecule-1 (sICAM-1) in patients with Helicobacter pylori Infection

ＩＮＴＲＯＤＵＣＴＩＯＮＳｉｎｃｅｔｈｅｆｉｒｓｔｓｕｃｃｅｓｓｆｕｌｉｓｏｌａｔｉｏｎｏｆＨ .ｐｙ ｌｏｒｉｉｎ 1 982 ,Ｈ .ｐｙｌｏｒｉｉｎｆｅｃｔｉｏｎｈａｓｂｅｅｎｆｏｕｎｄｔｏｂｅａｓｓｏｃｉａｔｅｄｗｉｔｈｖａｒｉｏｕｓｄｉｓｅａｓｅｓｉｎｃｌｕｄｉｎｇｃｈｒｏｎｉｃｇａｓｔｒｉｔｉｓ,ｐｅｐｔｉｃｕｌｃｅｒｄｉｓｅａｓｅｓ(Ｄｏｏｌｅｙｅｔａｌ.,1 989;Ｇｒａｈａｍｅｔａｌ.,1 992 ) ,ａｎｄｇａｓｔｒｉｃｎｅｏｐｌａｓｍｓ(Ｂａｙｅｒｄｏｒｆｆｅｒｅｔａｌ.,1 995) .Ｉｔｗａｓｓｕｇｇｅｓｔ…     

萌发花生种子中CaM含量变化与种子活力

应用ELISA测定了萌发花生种子中CaM的含量,种子的大部分CaM存在于胚轴中,小部分存在于子叶。高活力种子CaM含量较高,在吸胀48h出现一个特征峰,随种子活力下降,CaM含量也减少,其特征峰延迟至 72h才出现,且峰值也低,Ca~(++)可提高种子活力,也提高CaM的含量,使特征峰从72h提早到48h出现,TFP使种子CaM含量下降,直至萌发96h仍来山现特征峰,Ca~(++)、CaM与种子活力密切相关。     

Preparation of high-affinity rabbit monoclonal antibodies for ciprofloxacin and development of an indirect competitive ELISA for residues in milk

A convenient competitive enzyme-linked immunosorbent assay (ELISA) for ciprofloxacin (CPFX) was developed by using rabbit monoclonal antibodies (RabMAbs) against a hapten-protein conjugate of CPFX-bovine serum albumin (BSA). The indirect competitive ELISA of CPFX had a concentration at 50% inhibition (IC₅₀) of 1.47 ng/ml and a limit of detection (LOD) of 0.095 ng/ml. The mAb exhibited some cross-reactivity, however, not so high with enrofloxacin (28.8%), ofloxacin (13.1%), norfloxacin (11.0%), fleroxacin (22.6%), and pefloxacin (20.4%). And it showed almost no cross-reactivity with other antibiotics or sulfonamides evaluated in this study. The competitive ELISA kit developed here could be used as a screening tool to detect and control illegal addition of CPFX in food products. This kit had been applied to milk detection and the recovery rates from samples spiked by CPFX were in a range of 63.02%–84.60%, with coefficients of variation of less than 12.2%.     

用高度特异性王浆主蛋白1多克隆抗体ELISA法检测蜂王浆新鲜度(英文)

目的:为蜂王浆主蛋白1(MRJP1)的快速检测和鉴别提供科学依据,为蜂王浆的质量控制提供技术支持。创新点:首次比较了MRJP1特异性多克隆抗体与MRJP1重组表达蛋白多克隆抗体对王浆主蛋白家族的免疫反应差异,验证了蜂王浆中MRJP1蛋白降解与保温时间的相关性,建立了以MRJP1作为蜂王浆新鲜度生物标志物的快速检测方法。方法:通过蜂王浆主蛋白家族蛋白的氨基酸序列同源性分析,筛选出MRJP1的特异性多肽区域,进行人工合成,免疫兔子后取血清制备成特异性多克隆抗体。用蛋白质印迹法(Western blot)检测了MRJP1特异性多克隆抗体与MRJP1重组表达蛋白多克隆抗体对王浆主蛋白家族的免疫反应。以新鲜蜂王浆为对照品,用MRJP1特异性抗体酶联接免疫吸附剂测定(ELISA)法和变性电泳胶灰度扫描法分别测定保温(40°C)7~49天的蜂王浆中MRJP1含量的变化,并进行了相关性分析。结论:MRJP1的特异性抗体对MRJP1蛋白具有专一的免疫识别特性,可特异性地检测代表蜂王浆新鲜度的MRJP1含量变化,并鉴别蜂王浆的真伪。     

Study on the serum levels of soluble intercellular adhesion molecule-1 (sICAM-1) in patients with Helicobacter pylori Infection

Objective: To evaluate the interaction between serum levels of soluble intercellular adhesion molecule-1 (sICAM-1) and Helicobacter pylori (H. pylori) infection in patients with chronic gastritis and peptic ulcer. Methods: The serum levels of sICAM-1 in 205 patients with chronic gastric diseases were detected by ELISA method and the status of H. pylori was determined by histologic examination, RUT, 14C - UBT, and serology. The sera obtained from 18 healthy volunteers served as controls. Results: The serum levels of sICAM-1 were significantly higher in patients with H. pylori positive than those of H. pylori negative (889.43±32.52 ng/ml vs. 747.07±30.45 ng/ml, P<0.05). The serum levels of sICAM-1 in patients with mild, moderate and severe infection of H. pylori were 841.68±72.36 ng/ml, 905.43±37.59 ng/ml and 1012.54±49.34 ng/ml,respectively (P<0.05). The serum levels of sICAM-1 proved to be significantly correlated with the density of H. pylori colonization in gastric mucosa (rs =0.316, P<0.001). The serum levels of sICAM-1 in patients with chronic gastritis and peptic ulcer were significantly higher than those in healthy controls (P<0.05). Conclusions: These results indicated that H. pylori infection up-regulates the expression of sICAM-1.     

Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp简

A novel monoclonal antibody (MAb) against oxytetracycline (OTC) was generated and characterized. The MAb was used in the development of an enzyme-linked immunosorbant assay (ELISA)-based detection system. An OTC-bovine serum albumin (BSA) conjugate was prepared and used in the immunization of mice. A conventional somatic cell fusion technique was used to generate MAb-secreting hybridomas denoted 2–4F, 7–3G, and 11–11A. An indirect competitive ELISA (icELISA) was applied to measure the sensitivity and specificity of each MAb in terms of its 50% inhibitory concentration (IC50) and percentage of cross-reactivity, respectively. MAb 2–4F exhibited the highest sensitivity, with an IC₅₀ of 7.01 ng/ml. This MAb showed strong cross-reactivity to rolitetracycline, but no cross-reactivity to other unrelated antibiotics. When MAb 2–4F was used to detect OTC from shrimp samples, the recoveries were in the range of 82%–118% for an intra-assay and 96%–113% for an inter-assay. The coefficients of variation of the assays were 3.9%–13.9% and 5.5%–14.9%, respectively.     

Study on the serum levels of soluble intercellular adhesion molecule-1 (sICAM-1) in patients withHelicobacter pylori infection

Objective: To evaluate the interaction between serum levels of soluble intercellular adhesion molecule-1 (sICAM-1) andHelicobacter pylori (H. pylori) infection in patients with chronic gastritis and peptic ulcer. Methods: The serum levels of sICAM-1 in 205 patients with chronic gastric diseases were detected by ELISA method and the status ofH. pylori was determined by histologic examination, RUT,¹⁴C-UBT, and serology. The sera obtained from 18 healthy volunteers served as controls. Results: The serum levels of sICAM-1 were significantly higher in patients withH. pylori positive than those of H. pylori negative (889.43±32.52 ng/ml vs. 747.07±30.45 ng/ml,P<0.05). The serum levels of sICAM-1 in patients with mild, moderate and severe infection ofH. pylori were 841.68±72.36 ng/ml, 905.43±37.59 ng/ml and 1012.54±49.34 ng/ml, respectively (P<0.05). The serum levels of sICAM-1, proved to be significantly correlated with the density ofH. pylori colonization in gastric mucosa (r _s=0.316,P<0.001). The serum levels of sICAM-1 in patients with chronic gastritis and peptic ulcer were significantly higher than those in healthy controls (P<0.05). Conclusions: These results indicated thatH. pylori infection up-regulates the expression of sICAM-1.     

依沙佐米治疗难治性/复发性多发性骨髓瘤临床试验效果的Meta分析

目的:评价依沙佐米治疗难治性/复发性多发性骨髓瘤(RRMM)的疗效及安全性。方法:计算机检索依沙佐米治疗RRMM的临床试验,手工检索纳入查询到的所有文献,提取数据后,使用R软件3.5.0进行Meta分析。结果:共纳入8篇文献,包括1096例RRMM患者,含依沙佐米三联方案的疗效优于单联方案或双联方案,以依沙佐米联合来那度胺和地塞米松(IRD)方案疗效最佳;IRD方案腹泻,恶心,皮疹,血小板减少,呕吐,腹泻≥3级,低血压≥3级,恶心≥3级高于来那度胺和地塞米松(RD)方案,而失眠,贫血≥3级低于RD方案,差异有统计学意义(P<0.05)。结论:含依沙佐米三联方案治疗RRMM的疗效较依沙佐米单联方案或二联方案显著。IRD方案疗效最佳,其不良反应较RD方案增多。