狂犬病抗体免疫金标检测试纸的研制

目的应用酶联免疫原理和胶体金层析技术,研制狂犬病抗体免疫金标检测试纸。方法采用特殊的生产工艺,在玻璃纤维膜包被胶体金标记狂犬病抗原,在硝酸纤维素膜上检测线和对照线处分别包被狂犬病抗原和兔抗狂犬病抗体,制成狂犬病抗体免疫金标检测试纸(人用)。结果当待检样品阳性时,在检测线处形成抗原抗体的免疫复合物而凝聚显色;当待检样品阴性时,检测线处不形成抗原抗体免疫复合物不显色。整个试验过程只需15 m in。试纸与ELISA试剂比较,两者都具有微量、特异、准确的优点,且金标试纸独具操作方便、快速和结果直观、容易判定的优点。结论应用胶体金免疫层析技术建立的"狂犬病抗体免疫金标检测试纸",可以准确地检测出被检样品是否带有狂犬病抗体,同时还可以通过检测线颜色的深浅判定抗体的含量高低

Pollution Characteristics and Ecological Risk of PBDEs in Water and Sediment from an Electronic Waste Dismantling Area in Taizhou

以台州某电子垃圾拆解工业园为圆心,在半径为16 km的范围内,由近及远设计了C(3 km)、S(5~10 km)和R(10~16km)三圈共30个采样点,研究了该区域水及沉积物中多溴联苯醚(PBDEs)的污染特征与生态风险.结果表明,水中PBDEs含量为9.4~57.2 ng·L~(-1),平均值为25.9 ng·L~(-1);沉积物中PBDEs含量为3.7~38 775 ng·g~(-1),平均值为2 779 ng·g~(-1);BDE-209均为主要成分.水及沉积物中PBDEs含量的空间分布态势均为:C圈>S圈>R圈,沉积物中PBDEs含量和离工业园区中心的距离呈极显著负相关(P S > R. Furthermore,the concentrations of PBDEs in sediments showed significant negative correlation against the distance from the industrial park( P < 0. 01). Compared with other regions around the world,the PBDEs contamination was more serious in the area,which indicated that e-waste dismantling activity was one of the significant sources for PBDEs pollution. It was estimated that a total of 30. 7 t PBDEs( including 28. 9 t BDE-209) was discharged into surrounding environment as a result of dismantling industrial activities in last 40 years. A preliminary ecological risk assessment for PBDEs in water and sediments was conducted by hazard quotient method. The results demonstrated that the Penta-BDEs in the center of e-waste dismantling area( a radius of 1. 5 km) was at particularly high risk level and could cause serious influence on the ecological safety and human health.环境保护公益性行业科研专项(201309047

Neurotoxic Effects of Co-exposure to Lead and Dechlorane Plus on Zebrafish (Danio rerio) Embryos

以斑马鱼(Danio rerio)为研究对象,探讨铅(Pb); 、得克隆(DP)及二者联合急性暴露对斑马鱼胚胎的神经毒性作用。结果表明,Pb(5、20 mug·L~(-1))和DP(15、60; mug·L~(-1))单独暴露均会引起斑马鱼自主运动频率增加,触摸反应能力和自由游泳活力下降,并且抑制初级运动神经元的生长,加剧尾部细胞凋亡。但; 与20 mug·L~(-1) Pb单独暴露相比,高剂量联合暴露(20 mug·L~(-1) Pb + 60 mug·L~(-1); DP)使斑马鱼的自主运动频率显著降低(P < 0.05),触摸反应能力和自由游泳活力显著增强(P <; 0.05),初级运动神经元轴突长度显著增加(P < 0.05),尾部细胞凋亡减少。与5 mug·L~(-1); Pb单独暴露相比,低剂量联合暴露(5 mug·L~(-1) Pb + 15 mug·L~(-1) DP)也显著减少斑马鱼尾部的细胞凋亡(P <; 0.05); 。上述结果表明,Pb或DP单独暴露对斑马鱼均可引起神经毒性作用;但二者联合暴露对斑马鱼自主运动、触摸反应以及自由游泳活力的影响则表现为拮抗作用。Neurotoxic effects of acute exposure to lead (Pb) or Dechlorane Plus; (DP),or both were investigated using zebrafish (Danio rerio) embryos.; Results showed that exposure to Pb (5,20 mug·L~(-1)) or DP (15,60 mug·; L-1) alone increased spontaneous movement,decreased touch response and; free-swimming activity,inhibited axonal growth of primary motoneuron and; induced cell apoptosis in zebrafish embryos. Co-exposure to 20; mug·L~(-1) Pb and 60 mug·L~(-1) DP significantly decreased spontaneous; movement (P < 0.05),enhanced touch response and free-swimming activity; (P < 0.05),increased axonal length of primary motoneuron (P < 0.05) and; reduced cell apoptosis in zebrafish when compared to 20 mug·L~(-1) Pb; exposure alone. Co-exposure to 5 mug·L~(-1) Pb and 15 mug·L~(-1) DP also; significantly decreased cell apoptosis on the tail region when compared; to 5 mug·L~(-1) Pb exposure alone (P < 0.05). These results demonstrated; that Pb or DP exposure alone could induce neurobehavioral toxicity in; zebrafish, but Pb and DP co-exposure had antagonistic effects on; spontaneous movements,touch response and free swimming activity.国家自然科学基金项目; 高等学校博士学科点专项科研基

猪瘟病毒EO蛋白的原核表达及其重组蛋白活性测定

通过重叠PCR合成猪瘟病毒EO基因,将该片段定向插入到pET-22b载体中,构建原核表达载体pET-22b／EO,转化大肠杆菌BL21 (DE3),IPTG诱导表达,比较不同诱导条件下的蛋白表达,确定其最佳表达条件。重组蛋白主要以包涵体的形式表达,Ni2+亲和层析柱纯化蛋白,逐步透析法复性。通过方阵试验确定包被抗原的最适工作浓度,为了测定EO蛋白的活性,本文初步建立了检测猪瘟血清抗体水平的间接 ELISA方法,为开发检测猪瘟抗体诊断试荆奠定基础

猪圆环病毒2型抗体免疫金标检测试纸的研制

应用酶联免疫原理和胶体金层析技术,采用特殊的生产工艺,在玻璃纤维膜包被胶体金标记PCV-2抗原,在硝酸纤维素膜上检测线和对照线处分别包被PCV-2抗原和兔抗PCV-2抗体,制成猪圆环病毒2型抗体免疫金标检测试纸。当待检样品阳性时,在检测线处形成抗原抗体的免疫复合物而凝聚显色;当待检样品阴性时,检测线处不形成抗原抗体免疫复合物不显色。整个试验过程只需15min。试纸与ELISA试剂比较,两者都具有微量、特异、准确的优点,且金标试纸独具操作方便、快速和结果直观、容易判定的优点

Study and Apply for Detection of PRRS Anti-body by ColloidalGgold Stripe

应用酶联免疫吸附原理和胶体金层析技术,在玻璃纤维和硝酸纤维素膜的检测线和对照线上分别喷上胶体金PRRSV,PRRSV和兔抗PRRS抗体,从而制作成猪PRRS抗体免疫金标试纸,用该试纸与PRRS-ELISA试剂盒分别对30份猪血清及血液样品进行抗体水平检测,结果完全一致。说明金标试纸法是一种微量、特异、简便和结果容易判定的新的检测方法,非常适用于基层兽医站和猪场,特别是进行现场检测和开展大规模疫情普查时使用。Based on the principle of immune - ELISA and colloidal gold immunochromatography, colloidal gold stripe was prepared and applied toto the detection of anti - PRRS antibody. Gold PRRSV, PRRSV and anti - PRRS antibody was coated on the test region and control zone respectively.Anti - PRRS antibody in 34 whole blood and sera sample was detected by the strip and PRRS - ELISA method. Result achieved from the methods mentioned above are in complete agreement. It shows that the method established in this paper bears the merit of micro analysis, specificity, conveniece and the result is easy to be tested. This method is applicable for the detection of anti - PRRS antibody in a wide area and can be applied to general survey of PRRS disease

肝内胆管结石合并肝胆管癌的诊断——附16例报告

目的　探讨肝内胆管结石合并肝胆管癌的诊断方法。　方法　回顾性分析1997 ~2003年经病理证实的16例肝胆管结石合并肝内胆管癌的临床资料,并对其诊断进行总结。　结果　术前诊断正确率为37. 5% (6 /16),术中和术后病理发现10例(62. 5% )。其中左肝内胆管癌10例(62. 5% ),右肝内胆管癌5例(31. 3% )。病理诊断为胆管周围侵润型12例(75% ),肿块形成型4例。　结论　对年龄较大、长期反复胆管炎发作的慢性肝内胆管结石患者,出现用常规方法难以控制的发热、持续性上腹部疼痛时,

Study and Apply of Colloidal Gold Strip in Detecting the Antibody of pseudorabies

应用免疫学原理与胶体金层析技术相结合 ,采用双抗原夹心法建立了检测猪伪狂犬病抗体水平的“猪伪狂犬病抗体免疫胶体金快速检测试纸法” ,用该法与美国Idexx公司的猪伪狂犬病ELISA试剂盒 ,同时对 1 6份猪血液或血清进行抗体水平检测 ,符合率达 1 0 0 %。同时使用金标法对 1 0 0份猪血液和对应血清进行检测 ,结果也相同。试验结果显示 ,金标法与ELISA一样 ,具有微量、特异、准确等优点 ,且操作简易 ,而金标法不需要任何仪器 ,检测时间短 ,结果直观 ,容易判定 ,适用于基层兽医站、养猪场使用和大面积猪伪狂犬病抗体普查。Based on the principle of colloidal gold immunochromatography, a test strip was prepared and applied in detecting the antibody of pseudorabies. The test result was coincidence with ELISA made in USA Indexx cor. in examination of 80 blood or sera samples from pig、chicken、 duck、 cattle and goat. It shows that this strip has the same merits as that of ELISA test in microanalysis, high specialty and accurate. No special instrument is needed and the test can be completed easily in a short time. So it can be used widely in common veterinary services, hoggery and the general survey of pseudorabies disease

海洋平台的健康监测技术与安全评估研究

本研究针对海洋平台的安全监测开展了成套技术研发。本报告主要记载了以下研究内容的研究过程、方法和结果： 1）基于Bragg光纤光栅（FBG）的高精度、长寿命、高可靠性海洋平台的结构健康（安全）监测传感系统，包括应变、加速度、位移、温度等系列光纤传感器和水下传感网络的密封、机械保护等； 2）结构整体、构件和局部热点部位多层次损伤反演理论和技术，包括基于FBG的应变模态损伤识别技术、相空间法损伤识别技术和局部热点损伤重构及成像技术； 3）结构安全状况和残余寿命的动态评估方法； 4）基于人工智能技术的结构安全评估专家系统； 5）海洋平台健康监测软硬件技术系统集成。 本技术可实现海洋平台的全天候、全方位自动化安全监测，显著提高结构的安全保障水平；同时可实现基于安全状况的海洋平台维修策略和方案，从而节约安全管理成本；可准确把握海洋平台的实际寿命，延长安全服役期

猪圆环病毒2型ORF1和ORF2基因的克隆与表达

根据GenBank猪圆环病毒2型基因序列,设计合成两对特异性引物,对厦门株-1进行PCR扩增,得到ORF1和ORF2基因。将PCR产物酶切后,插入到pET-22b载体中,构建原核表达载体pET-22b/ORF1、pET-22b/ORF2,转化大肠杆菌BL21(DE_3),经IPTG诱导表达,收集茵液进行SDS-PAGE分析。确定重组蛋白主要以包涵体的形式表达,包涵体洗涤溶解后,采用Ni~(2+)离子金属螯合亲和层析柱纯化蛋白,逐步透析法进行复性。Western blot和ELISA分析结果表明Rep蛋白和Cap蛋白均能与猪圆环病毒2型阳性血清发生特异性反应