海产贝类帕金虫病害的研究进展

比较系统地介绍了国外贝类帕金虫病害的研究进展，包括帕金虫的分类地位、形态结构、生活史、检测方法、病理学、病害发生生态机理、防治技术与对策等。我国目前海产贝类病害越来越重，并且在发病贝类体内也检测到了帕金虫的存在。因此，系统阐述国外贝类帕金虫病害的研究进展，对我国海产贝类病害的研究，具有现实意义。     

氯菊酯农药间接酶联免疫吸附测定法的建立

为测定环境样品中氯菊酯农药残留,用活泼酯法将氯菊酯半抗原(Py)与卵白蛋白(OVA)偶联,制备合成抗原Py-OVA作包被原,建立间接竞争酶联免疫吸附测定法.方阵滴定确定了抗血清最佳稀释度(1∶2 500),包被抗原的最适工作浓度0.45μg/mL,并建立了标准工作曲线.工作曲线表明在10～800μg/L浓度范围内呈良好的线性关系,回收率>97%.     

温度盐度对菲律宾蛤仔帕金虫病害发生的影响

在实验室内进行了暖冬、海水温度和盐度对菲律宾蛤仔体内寄生帕金虫数量变化影响的实验,结果表明,在冬季高于自然海水温度的各实验组中菲律宾蛤仔体内的帕金虫含量要高于自然海水温度实验组,并且显示出菲律宾蛤仔体内过冬的帕金虫于春末和夏初当海水温度达到20℃左右时又出现明显增高趋势,导致菲律宾蛤仔的死亡率显著提高;盐度实验结果表明,在温度相同情况下,20～24的盐度是菲律宾蛤仔体内帕金虫的最适繁殖盐度.针对实验结果,提出了预防菲律宾蛤仔的帕金虫病害的一些生态养殖措施.     

菊酯类农药酶联免疫吸附测定研究进展

介绍了菊酯类农药残留的酶联免疫分析(ELISA)的技术原理、关键技术环节,综述了国内外的酶联免疫测定菊酯类农药残留的研究动态,并展望了此技术的发展及应用前景。     

单克隆抗体间接竞争ELISA测定水体中氟虫腈

制备了氟虫腈单克隆抗体 ,在此基础上建立了水体中氟虫腈测定的间接竞争性酶联免疫吸附测定 (ELISA)法 .在优化条件下 ,氟虫腈测定的线性浓度范围为 1 0 -1 μg·L-1 ～ 1 0 3μg·L-1 ,最低检出浓度 (I1 0 )为 0 0 8μg·L-1 ,检测灵敏度 (I50 )为6 6 8μg·L-1 ;常见的几种与氟虫腈结构类似的农药的交叉反应率均低于 1 % ,不干扰氟虫腈的检测 ;2 0 %丙酮、5 %甲醇、2 %二甲基甲酰胺、2 %乙腈和 2 %乙酸乙酯基本不影响免疫测定 .利用建立的间接竞争性ELISA方法检测不同水样中氟虫腈 ,结果表明不同水样均可直接进行测定 ,方法的精密度和准确度均符合残留测定的要求     

快速检测软骨藻酸的间接ELISA方法

分别对抗原、一抗、二抗的最佳稀释倍数、温育温度与时间、包被条件及其显色条件进行了优化,建立了一种快速检测软骨藻酸(Domoic acid,DA)的酶联免疫分析法(ELISA).结果表明:抗原、一抗和二抗(HRP-IgG)最佳稀释度分别为1∶12800倍、1∶400倍和1∶3000倍;最佳包被条件为4℃包被12 h;抗原抗体最佳反应温度为37℃,反应时间为60 min;二抗最佳反应温度为43℃,反应时间为30 min;最佳显色条件为室温不避光显色20 min.该方法的样品加标回收率高达86.8%～103.2%,其最低检测限(LOD)为4.86 ng·mL-1,光吸收值(OD值)的变异系数(CV)在2.46%～7.08%之间.     

恩诺沙星完全抗原的合成及间接竞争ELISA方法的建立

为了建立环境新型抗生素类污染物恩诺沙星的免疫检测方法,采用间接竞争ELISA方法,用酶标仪测定酶标二抗与底物反应生成的有色产物量来确定恩诺沙星含量.研究了恩诺沙星完全抗原的合成方法,以及间接竞争ELISA的优化条件和实际水样的检测效果.结果表明,通过碳二亚胺法(EDC+NHS)成功合成了恩诺沙星的理想完全抗原,采用BCA法测定完全抗原浓度,紫外吸收法和MADLI-TOF MS法测得完全抗原的偶联比为14~16.建立的恩诺沙星间接竞争ELISA检测方法,检测限为1.92μg·L~(-1),定量检测区间为7.33~238.24μg·L~(-1).采用优化后的实验条件测定了清华大学地下水和饮用水配制的恩诺沙星加标样品,发现所有样品的变异系数和回收率分别为3%~11%和82%~122%,表明建立的间接竞争ELISA方法适用于实际水样中恩诺沙星的快速简便检测.     

水体中微囊藻毒素-LR的间接竞争ELISA检测

在自制包被完全抗原浓度为5μg/mL、单克隆抗体工作稀释度为1∶3 000、酶标二抗鼠工作稀释度为1∶3 000、微囊藻毒素-LR浓度在0.001～30μg/L、显色底物为邻苯二胺,采用间接竞争酶联免疫吸附试验对水体中的微囊藻毒素-LR进行检测,结果表明,该方法与高效液相色谱的检测结果相关系数大于0.99,多次重复实验相对标准偏差小于10%,最低检测限能达到0.01μg/L,定量检测区间为0.01～3μg/L,该方法对[4-精氨酸]微囊藻毒素能特异性识别,对来自实际水样中的干扰有相当的耐受力.     

酶联免疫吸附分析法测定水样中的毒死蜱

自制了毒死蜱多克隆抗体,考察了离子强度和甲醇含量对抗原抗体竞争反应的影响,对反应体系进行了优化,最终建立了水样中毒死蜱的残留检测ELISA方法。结果表明,毒死蜱对抗体竞争性结合的I50为76.8μg/L,方法检出限为5.2μg/L。河水样品经简单前处理后分别采用ELISA和GC测定,用ELISA方法测得水样中毒死蜱的添加回收率为93.40%～102.1%,C.V为4.04%～5.18%,在水样中的最低检出浓度为0.08μg/L,检测灵敏度远高于GC,符合农药残留检测的要求。     

小麦中T-2毒素酶联免疫吸附测定法(ELISA)

T-2毒素是某些植物致病菌的次生代谢产物,尤其是由镰刀菌(Fusarium sp)引致的麦类、玉米等作物的赤霉病,菌体在一定条件下能产生大量的该类毒素.T-2毒素对人畜的毒性很大,同时,还与某些癌症的发病率有密切的关系.T-2毒素的检测方法,常用的有薄层层析法,气相、液相色谱法以及气-质联用等,这些方法,有的精确度     

Microcystin-LR detection based on indirect competitive enzyme-linked immunosorbent assay

Microcystins (MCs) are a group of closely related toxic cyclic heptapeptides produced by common cyanobacteria, which cause lots of accidents and threatens human health. In this paper, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was established and used to detect microcystin-LR (MC-LR) in drinking and surface waters. The concentration of coating antigen was 5 μ/mL, the dilution of monoclonal antibody MC10E7 was 1:3 000, the dilution of enzyme tracer (goat anti-mouse IgG-peroxidase) was 1:3 000, the standard concentration of MC-LR ranged from 0.001 μg/L to 30 μg/L, and o-phenylenediamine was used as substrate. The assay showed high relativity with high performance liquid chromatography (HPLC) with a correlation coefficient of more than 99%. The relative standard deviation was less than 10%, the detection limit was achieved down to 0.01 μg/L and up to 5.1 μg/L. The quantitative detection range was from 0.03 μg/L to 3 μg/L, and the antibody had high specificity for [4-arginine] microcystins. It performed well in spite of the influence of the real samples. Translated from Environmental Science, 2006, 27(6): 1166–1170 [译自: 环境科学]     

Determnation of ochratoxin A in grain by monoclonal antibody-based enzyme-linked immunosorbent assay

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酶联免疫吸附法测定水中异丙隆

建立了测定水中异丙隆的酶联免疫吸附法。灵敏度0.97μg/L，最低检出限0.03μg/L16种取代脲类除草剂的交叉反应率低于2.2%。水样可不经分离提取直接测定。10%甲醇、2%乙腈和2%乙醇不影响测定。比较了11种与异丙隆结构相近的半抗原的活性。     

间接竞争ELISA法测定稻田土壤中除草剂毒莠定的残留量

通过对反应体系中酶标二抗(辣根过氧化物酶标记羊抗兔IgG)和抗体的最佳稀释浓度等条件的筛选和确定,最终建立了一种快速灵敏地测定污染稻田土壤中毒莠定残留量的方法--间接竞争ELISA分析方法.结果表明,酶标二抗和抗体的最佳稀释度均为1:500(体积比),毒莠定的检出下限达到5 ng·mL-1,在0.07～0.7μg·mL-1浓度范围内有良好的线性;土壤浸出液加样的平均回收率为104.11%,变异系数在3.13%～11.13%之间,符合农药残留分析的要求;样品基质对检测结果没有干扰.     

间接免疫荧光抗体技术检测凡纳滨对虾红体病病原——副溶血弧菌

建立了凡纳滨对虾红体病病原--副溶血弧菌的间接免疫荧光抗体检测方法.确定免疫血清和荧光抗体的最适工作浓度分别为1∶ 1000和1∶ 200;交叉反应和阻断试验表明该方法具有较高的特异性.用该方法检测了10份人工感染后发病的凡纳滨对虾和10份感染后外观健康的凡纳滨对虾,阳性检出率分别为100%和60%,表明该技术不仅能够检测已发病的凡纳滨对虾,而且能够检测带菌的凡纳滨对虾,这对于水产养殖业中疾病的早期诊断有着重要意义,而且该方法具有快速、特异、简单的优点,适合于现场应用推广.     

Enhanced the photoelectrochemical performance of Bi2XO6 (X = W, Mo) for detecting hexavalent chromium by modification of CuS

In this work, Bi₂XO₆ (X = W, Mo) are synthesized at different temperatures. The results of tests find the optimal temperatures of Bi₂WO₆ and Bi₂MoO₆ are 180 and 160°C (BW-180, BM-160). Then, BW-180 and BM-160 are further compounded with different contents of CuS. The results of photoelectrochemical (PEC) tests show that CuS can improve the PEC performance of semiconductor materials, and it has better performance when CuS mass fraction is 5%. These maybe the photoelectron potentials generated by CuS/Bi₂XO₆ (X = Mo, W) heterojunction reduce the combination of photogenerated electrons and holes. When the PEC sensor based on 5%-CuS/BW-180 detects Cr(VI), it has a linear range of 1–80 μmol/L with detection limit of 0.95 μmol/L, while the PEC sensor based on 5%-CuS/BM-160 detects Cr(VI) has a linear range of 0.5–230 μmol/L and a detection limit of 0.12 μmol/L. Thus, 5%-CuS/Bi₂XO₆ has potential application in hexavalent chromium detection.     

Expression and Immunoreactivities of Hepatitis E Virus Genotype 3 Open Reading Frame-2 (ORF-2) Recombinant Proteins Expressed in Insect Cells

Hepatitis E virus (HEV) is a fecal-orally transmitted virus that is endemic in many geographical areas with poor sanitary conditions and inadequate water supplies. In Europe, a low-endemic area, an increased number of autochthonous sporadic human cases of patients infected with HEV strains of genotype 3, have been reported lately. The relatively high prevalence of HEV genotype 3 infections in European pigs has raised concerns about a potential zoonotic transmission to humans. Determination of HEV seroprevalence in pigs would help to clarify its incidence and possible zoonotic implications. To this purpose, we have expressed and partially characterized swine genotype 3 HEV open reading frame-2 proteins upon infection of Sf21 insect cells with recombinant baculoviruses. The use of the expressed proteins as diagnostic antigens for the detection of antibodies to HEV has been further assayed with human and swine sera.