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581.
【背景】非洲猪瘟(African swine fever, ASF)作为高致病性传染病,给我国生猪养殖业造成了严重的经济损失,因此建立快捷、灵敏的诊断方法至关重要。【目的】建立一种管式非洲猪瘟病毒pp62蛋白化学发光抗体检测方法。【方法】以重组pp62蛋白作为包被抗原,羧基磁珠(carboxylic magnetic beads)作为固相载体,碱性磷酸酶(alkaline phosphatase, AP)标记的兔抗猪IgG作为酶标二抗,通过对反应条件进行优化,采用非洲猪瘟国家参考品作为溯源血清绘制出标准曲线,建立基于ASFV pp62蛋白的化学发光抗体检测方法。【结果】用建立的化学发光方法检测277份临床样品血清,通过受试者工作特征(receiver operating characteristic, ROC)曲线分析确定阴性、阳性临界值,并确定判定标准:浓度值>140.02 U时为抗体阳性,浓度值<140.02 U时为抗体阴性。此方法对6种不同的病原血清抗体进行检测均无交叉反应,且批内和批间变异系数均在10%以内,与商品化非洲猪瘟抗体检测试剂盒的符合率达96.7%。【结论】本研究建立的管式非洲猪瘟病毒化学发光抗体检测方法具有良好的特异性、敏感性和重复性,可为非洲猪瘟早期监测及试剂盒研发提供参考。 相似文献
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猪瘟病毒衣壳蛋白靶向核酸酶表达系统的建立及鉴定 总被引:1,自引:0,他引:1
根据猪瘟病毒衣壳蛋白(C)基因序列设计一对引物,RT-PCR扩增获得编码猪瘟病毒衣壳蛋白的C基因,将其插入到含有葡萄球菌核酸酶(SN)基因的真核表达载体pcDNA-SN中,筛选获得重组质粒pcDNA-C-SN。脂质体转染猪肾细胞(PK-15),并经G418稳定筛选,通过RT-PCR、免疫印迹和间接免疫荧光鉴定表达的融合蛋白,体外DNA消化试验检测核酸酶活性。结果表明融合蛋白C-SN在PK-15细胞中获得了稳定表达,能够被兔抗猪瘟病毒衣壳蛋白多抗所识别,并具有良好的核酸酶活性,能够对DNA进行切割。同时,稳定表达融合蛋白C-SN的PK-15细胞系中能够有效抑制猪瘟野毒的增殖,使其感染性降低102~103倍。这些结果为进一步将衣壳蛋白靶向病毒灭活策略应用于抵抗猪瘟病毒感染奠定了基础。 相似文献
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JAMES A. COMER WILLIAM S. IRBY DARRELL M. KAVANAUGH 《Medical and veterinary entomology》1994,8(4):325-330
Abstract. Hosts of Lutzomyia shannoni Dyar, a suspected biological vector of the New Jersey serotype of vesicular stomatitis (VSNJ) virus, were determined using an indirect enzyme-linked immunosorbent assay (ELISA) of 333 blood-fed female sandflies collected from their diurnal resting shelters on Ossabaw Island, Georgia, U.S.A. Sandflies had fed primarily on white-tailed deer ( Odocoileus virginianus ) (81%) and to a lesser extent on feral swine ( Sus scrofa ) (16%), two species of host infected annually with VSNJ. Other hosts were raccoons ( Procyon lotor ) and horses ( Equus caballus ) or donkeys ( E. asinus ), with only two (<1%) mixed bloodmeals from deer/raccoon and deer/swine. A larger proportion of feedings on feral swine was detected in maritime live oak forests than in mixed hardwood forests. These findings are consistent with the hypothesis that L. shannoni is a primary vector of VSNJ virus on Ossabaw Island. 相似文献
586.
《Journal of Asia》2020,23(4):1138-1143
Edible insects are increasingly recommended as novel sustainable protein sources, but the nutritional properties of edible insects have not been well studied. We investigated whether locust powder can be used as a nutritionally functional food resource with a low energy value using a rat model. Twenty-five male Wistar rats (4-week-old) were fed a basal diet (12 g daily) to which a fixed amount of locust powder (0, 0.5, 1.0, or 2.0 g) was added for 20 days (L0, L0.5, L1.0, and L2.0 groups, respectively). In the sucrose standard group, rats received 12 g of the basal diet daily and 2.0 g of sucrose daily for 20 days (S2.0 group). Body-weight gain and the nutritional composition of the carcasses and feces were determined to estimate the available energy value of locust powder. The L0.5 group had the lowest carcass fat content and energy accumulation, but these values were increased by locust powder in a dose-dependent manner. The net energy value of locust powder was estimated to be 2.78 kcal/g, which was expected to be lower than the calculated theoretical value (4.25 kcal/g) and that of sucrose (3.94 kcal/g). Fecal nitrogen excretion was increased by dietary locust powder in a dose-dependent manner (correlation coefficient, R = 0.98), and the carcass nitrogen percentage was not changed, regardless of the dietary content of the locust powder, indicating an increased excretion of proteins or other non-protein nitrogen compounds derived from the locust powder. These findings suggest that locust powder can be used as a novel food material with a low energy value for humans. 相似文献
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Yu-Bei Jin Wen-Tao Yang Ke-Yan Huang Hong-Liang Chen Seria-Masole Shonyela Jing Liu 《Bioscience, biotechnology, and biochemistry》2017,81(8):1489-1496
Recombination activating gene 2 (RAG2) is necessary for immature B cell differentiation. Antibodies to human and rabbit RAG2 are currently commercially available, but antibodies to swine RAG remain unavailable to date. In this study, the swine RAG2 genes sequence was synthesized and then cloned into a pET-28a vector. The recombinant fusion protein was successfully expressed in E. coli, purified through nickel column chromatography, and further digested with Tobacco Etch Virus protease. The cleaved protein was purified by molecular-exclusion chromatography and named pRAG2. We used pRAG2 to immunize rabbits, collected the serum and purified rabbit anti-pRAG2 polyclonal antibodies. The rabbit anti-pRAG2 polyclonal antibodies were tested via immunofluorescence on eukaryotic cells overexpressing pRAG2 and also able to recognize pig natural RAG2 and human RAG2 protein in western blotting. These results indicated that the prepared rabbit anti-pRAG2 polyclonal antibodies may serve as a tool to detect immature B cell differentiation of swine. 相似文献