基于量子点的赭曲霉毒素AsFLISA检测技术研究

建立了基于量子点标记技术的赭曲霉毒素A(Ochratoxin A,OTA)双抗夹心荧光免疫检测(sandwich fluorescence-linked immunosorbent assay,sFLISA)体系,包含多克隆包被抗体、生物素标记的多克隆检测抗体、量子点标记的链霉亲和素等,获得了sFLISA的最佳操作参数:包被抗体浓度2.5μg/mL,检测抗体稀释500倍,量子点标记链霉亲和素稀释100倍。该方法在OTA浓度3.125~125μg/L之间时,相对荧光强度和OTA浓度呈线性关系,回归方程为y=0.0206x+0.2018,R2=0.9924;加标回收率在90.1%~110.0%之间,变异系数均小于10%,能较好地进行赭曲霉毒素A的定量检测。     

Screening of 170 Peruvian plant species for allelopathic activity by using the Sandwich Method

Peru is one of the 20 botanically extremely diverse countries in the world, with >17 000 flowering plants, of which 30% are endemic. So far, no systematic research has been conducted on the screening of the allelopathic plants. In this study, the allelopathic activity of 170 species from 61 families of Peruvian plants that were collected from the three main regions of Peru – the Costa (Pacific coastline), the Sierra (Andean mountains), and the Selva (Amazonian rainforest) – was evaluated. The allelopathic activity was determined by the Sandwich Method, which can evaluate the activity of leaf leachates. The species that were found to be highly inhibitory in this screening, under the criterion of >90% inhibition of the radicle of lettuce (Lactuca sativa) seedlings, were Aristeguietia ballii and Diplostephium foliosissimum (Asteraceae) and Spondias mombin (Anacardiaceae). All of these species are native plants from Peru. This study gives a strong clue regarding the potential of isolating potent allelochemicals from these plants in the future.     

Sandwich panel of veneer-overlaid low-density fiberboard

Low-density sandwich panels of veneer-overlaid fiberboards of 12 mm thickness for structural use were manufactured at densities of 0.3–0.5g/cm³ using an isocyanate compound resin adhesive and steam injection pressing method. The effects of board density, veneer thickness, and resin content on the fundamental properties of sandwich panels were examined, with the following results: (1) The dry moduli of rupture and elasticity in the parallel direction of sandwich panels with thicker veneers were superior. The dry moduli of rupture and elasticity in the parallel direction of sandwich panels with 2.0 mm thick veneer at densities of 0.4–0.5 g/cm³ were 40–60 MPa, and 5–8 GPa, which were two and four times as much as those of homogeneous fiberboards, respectively. (2) The higher-density panels exhibited tensile failure at the bottom veneer surface during static dry bending in a parallel direction, whereas lower-density panels experienced horizontal shear failure in the core. (3) The dimensional stability of sandwich panels had good dimensional stability, with negligible springback after accelerated weathering conditions. (4) The thermal insulation properties of sandwich panels were found to be much superior to other commercial structural wood composite panels.Part of this report was presented at the 47th annual meeting of the Japan Wood Research Society, Kouchi, April 1997     

猪圆环病毒2型Cap蛋白ELISA检测方法的建立及初步应用

将杆状病毒表达系统表达并经梯度离心纯化的PCV2Cap蛋白作为标准品,利用双抗夹心法建立了针对猪圆环病毒2型Cap蛋白的ELISA检测方法,并应用于圆环病毒2型基因工程疫苗的质控。结果表明,多抗最适包被浓度为1μg/mL,最佳封闭液为1％BSA,最佳单抗反应浓度为2μg／mL,反应时间60min,最佳二抗使用稀释度1：40000,反应时间60min。该方法与St9细胞、BSA和其他猪病病毒抗原之间无交叉反应,最低检测抗原量为5ng。     

禽流感(H9N2亚型)酶标抗体的制备及双抗夹心Dot-ELISA检测方法的建立

采用AIV灭活油乳苗（H9N2）对AIV非免疫鸡进行接种并获取高免血清，取高免血清采用饱和硫酸铵沉淀法，经过Sephadex G25层析柱提纯后可获得纯度很好的IgG。将IgG应用过碘酸钠法标记辣根过氧化酶（HRP），制备禽流感酶标抗体（IgG—HRP），利用所提取的AI—IgG和所制备的禽流感酶标抗体按照双抗夹心Dot—ELISA试验操作步骤，采用方阵法分别摸索包被抗原及酶标抗体的最佳浓度，以及各步反应时间等最佳反应条件，以建立一种优化的AIV双抗夹心Dot—ELISA检测法。结果表明，所制备的AI高免血清HI效价为10log2；AI酶标抗体克分子比值为2．45；优化的Dot—ELISA各条件为：包被AI—IgG最佳稀释倍数为1：50；最佳封闭剂为1％牛血清白蛋白；AI酶标抗体最佳稀释倍数为1：100；待检抗原与2种抗体的感作时间均为0．5h（37℃）。优化后的检测方法可在2．5h内诊断结果。制备好的包被膜在4℃下保存2个月不影响其效果。而且敏感性提高了3倍，与新城疫病毒液、传染性支气管炎及减蛋综合症病毒液无交叉反应。     

林地面积空间抽样方法研究

论文采用空间简单随机抽样、空间分层抽样和三明治空间抽样模型,利用两种布样方式,随机和分层布样。通过对总体样本量和空间相关性的估算,进行样本分配和空间分层,结果表明：三明治空间抽样模型的抽样精度最高,其次是空间分层抽样、简单随机抽样;布样时采用分层抽样方式,将地理区域上分布的对象依照相似的属性值划分到不同的区域里,可以明显提高抽样调查效率及估计精度。     

魔芋加入量对猪耳西式火腿物性的影响研究

探讨了发酵猪耳西式火腿加工过程中魔芋加入量对产品物性的影响,旨在为火腿制品的开发提供一定的实验依据;为猪耳的利用拓展一条新的途径。结果表明:加入质量分数为83.3%的魔芋凝胶,可使制品获得较理想的感官性能;碱性魔芋精粉溶液的加入对产品质量有明显影响,其加入量以25.7%为宜。     

单克隆抗体ELISA双抗体夹心法与ELISA间接法检测小鼠肝炎病毒抗体的比较

用辣根过氧化物酶标记的抗小鼠肝炎病毒(MHV)单克隆抗体(McAb),建立了ELISA 双抗体夹心法,并与 ELISA 间接法检测小鼠血清中的 MHV 抗体进行了比较。发现两种方法在检出率、特异性、稳定性方面并无明显差异,但由于间接法操作更为简便,因此认为,检测血清中 MHV 抗体采用间接法较好。     

单抗夹心ELISA检测产蛋下降综合征病毒的研究

利用３株特异性单抗,建立了检测ＥＤＳＶ的夹心ＥＬＩＳＡ方法。经测定,ＭｃＡｂ最适包被浓度为２μｇ／ｍｌ,ＭｃＡｂ－ＨＲＰ最佳工作浓度为１∶４００,该方法最小检出浓度为０．０３μｇ／ｍｌ,与常见的几种鸡的传染病的病原无交叉反应。对某发病鸡场的５０份泄殖腔拭子样本进行了检测,阳性率为８４％（４２／５０）。     

猪伪狂犬病病毒单抗双夹心ELISA的构建

建立了检测猪伪狂犬病病毒(Pseudorabiesvirus,PRV)的单克隆抗体双夹心酶联免疫吸附试验(Enzyme linkedimmunosor bentassay,ELISA)方法,为该病的快速诊断奠定了基础.作者对其组装条件进行了筛选,认为兔抗PRVIgG最佳包被质量浓度为4.59mg·L-1;抗PRV的单克隆抗体(Monoclonalantibodies,McAb)的最佳质量浓度为6.45mg·L-1;最佳封闭剂为10g·L-1BSA(牛血清白蛋白),封闭时间为60min,制定了科学的结果判定标准,最低病毒检出量为200μg·L-1.与病毒分离、动物试验和中和试验相比较,该方法特异、灵敏、可靠、方便.