猪瘟病毒结构蛋白抗体间接ELISA检测方法的建立及评价     

袁莉  杨顺利  尚佑军  贾怀杰  景志忠  刘湘涛  蔡建平  尹双辉 《西北农业学报》2018,27(9):1273-1279

通过研究猪瘟(CSF)疫苗免疫猪抗结构蛋白E2、E0和C抗体的产生规律,为临床制定合理的CSF疫苗免疫程序提供参考。利用间接ELISA方法检测CSF疫苗免疫猪过程中,抗E2抗原血清抗体的产生特征;分别以重组猪瘟病毒(CSFV)结构蛋白E0和C为抗原,建立检测血清抗体的间接ELISA方法,评价其敏感性和特异性,并检测疫苗免疫过程中抗E0和C蛋白抗体的消长规律。结果显示:CSFV-E0和CSFV-C间接ELISA抗体检测方法的抗原最佳包被质量浓度分别为2.09和1.59μg/mL,待检血清样本最适稀释度分别为1∶60和1∶80,酶标二抗稀释度分别为1∶100和1∶350,2种ELISA方法与常见猪病原的阳性血清无交叉反应,具有良好的特异性。CSF疫苗免疫后第7天即可检测到抗E2、E0和C蛋白的抗体,其中抗E2和E0的抗体在免疫后第21天达到最高水平,E2抗体滴度先上升后下降,E0抗体在免疫后第21天开始下降并最终趋于稳定;C蛋白抗体在免疫后第7天出现,随后开始下降,在第28天转为阴性。结果表明,利用重组抗原建立的CSFV血清抗体ELISA检测方法,可以用于评价CSF疫苗免疫后血清中针对3种结构蛋白的抗体消长特点,从而为临床猪瘟疫苗免疫程序的制定提供参考。  相似文献   

大型溞ChE和NAGase间接竞争和非竞争ELISA方法的比较     

郎倩萍  李少南  郑佳豪  张文萍 《安徽农业科学》2016,(24):1-5

[目的]探索间接非竞争ELISA和间接竞争ELISA法测定大型溞ChE和NAGase的试验条件及方法灵敏度。[方法]分别采用间接竞争和间接非竞争ELISA方法测定大型溞ChE和NAGase。[结果]对于ChE,间接非竞争ELISA的最适抗原浓度和抗体稀释倍数分别为1.466μg/m L和1∶10 000,灵敏度为7.426 7 ng/m L;间接竞争ELISA的最适抗原浓度和抗体稀释倍数分别为5.277μg/m L和1∶8 000,灵敏度为0.163 4 ng/m L。对于NAGase,间接非竞争ELISA的最适抗原浓度和抗体稀释倍数分别为8.961μg/m L和1∶6 000,灵敏度为4.199 9 ng/m L;间接竞争ELISA的最适抗原浓度和抗体稀释倍数分别为6.450μg/m L和1∶4 000,灵敏度为0.250 8 ng/m L。[结论]无论是ChE还是NAGase,间接竞争ELISA在最适抗体浓度下的检测灵敏度均高于间接非竞争ELISA,加之其操作简便,间接竞争ELISA在生物和环境样品ChE和NAGase含量检测中值得推广应用。  相似文献   

Effects of sika deer on vegetation in Japan: A review     

Seiki Takatsuki 《Biological conservation》2009,142(9):1922-1929

The Japanese sika deer Cervus nippon has expanded its range by nearly 70% during the last two decades. Browsing by sika deer affects vegetation in both agricultural and forested habitats. Effects of sika deer on vegetation are conspicuous on deer-inhabited islands: forest structure and composition are altered by deer grazing and browsing, and consequently regeneration is prevented. By felling of old trees, forest gaps are formed, but since sapling recruitment is prevented, shade intolerant plants invade. Unpalatable forbs like ragwort Senecio cannabifolius and ferns like bracken Pteridium aquilinum (Dennstaedtiaceae) become dominants in open habitats. At the places of highest deer density, the Zoysia japonica community, a low growing grass mat, develops. Sika deer function as a seed dispersal agent of this grass. Indirect effects of sika deer are not well studied, but some studies have shown reduction of understory bamboo cover, which in turn improves the survival of tree seedlings and declines of wood mice Apodemus spp. Japanese forests in lower mountainous areas were widely logged during the 1940s and 1950s. These areas were not well planted during the war and until 1950, but thereafter intensively planted to alter to conifer plantation as a nationwide campaign. For several decades after the war, rodents and hares grazed planted trees. According to growth of the planted trees to form canopy, which is unfavorable for rodents and hares, their damage declined. After the 1960s, old-growth forests in high mountainous areas were logged, and conifers were planted. Animals causing forestry damage were replaced by sika deer during the 1980s. Sika deer eat a wide variety of foods, and are gregarious, which causes heavy impacts on vegetation. Effects of sika deer are expanding to natural forests, alpine vegetation, and marshes. To reduce damages on forestry and natural vegetation, as many as 100,000 sika deer are culled every year. However, the hunter population is rapidly declining, and it is expected population control by culling will be insufficient. Although sika deer are an important component of Japanese forests, current population densities exceed the capacity of many ecosystems to tolerate herbivory.  相似文献   

Development and application of G₁-ELISA for detection of antibodies against bovine ephemeral fever virus     

Fu-ying Zheng  Guo-zhen Lin  Chang-qing Qiu  Ji-zhang Zhou  Xiao-an Cao  Xiao-wei Gong 《Research in veterinary science》2009,87(2):211-212

The gene encoding antigenic site G₁ of bovine ephemeral fever virus (BEFV) was highly expressed in the host cell Escherichia coli. An indirect G₁-ELISA with the recombinant protein as the coating antigen was established to detect antibodies against BEFV. The result revealed that the optimal concentration of the coated antigen was 0.5 μg/well and the dilution of serum was 1:20. It was optimal that sera with P/N value  2.2 were considered positive, P/N value  2.0 negative, and between 2.0 and 2.2 ambiguous. The G₁-ELISA method gave a sensitivity of 97.6% and a specificity of 98.6% by testing 590 field serum samples. These results suggest that the G₁-ELISA may be a good alternative tool for seroepidemiological surveys.  相似文献   

蒜头果凝集素的提取及血凝效果研究   总被引：1，自引：1，他引：0  

袁燕袁再美  岑晓江程霞 《中国农学通报》2011,27(21):144-146

从蒜头果种仁中提取凝集素,用20%、40%、60%、80%、100%饱和度硫酸铵分级沉淀,分别用鸡、兔的红细胞悬液检测其凝血活性。结果表明,80%以下硫酸铵沉淀物均可与鸡、兔的红细胞悬液发生凝集反应,100%的硫酸铵沉淀物则没有凝集反应,说明用硫酸铵沉淀到80%饱和度以后,蒜头果凝集素即可完全沉淀出来。此结果证明蒜头果中含有凝集素,为蒜头果凝集素的开发利用奠定了基础。  相似文献   

口蹄疫病毒结构蛋白VP1上B细胞表位的筛选鉴定   总被引：1，自引：0，他引：1  

张昱  王永录  张永光  方玉珍  潘丽  蒋守田  吕建亮  刘力宽  张中旺  张淑刚  李正丰  杜进鑫 《华北农学报》2009,24(5)

以口蹄疫病毒株AF72 RNA为模板,反转录并扩增结构蛋白VP1基因,PCR纯化产物与pGEM-T easy载体连接并转化JM109菌株,对经凝胶电泳、PCR和EcoRⅠ酶切法鉴定为阳性的重组质粒进行测序,通过序列比对获得AF72 VP1的核苷酸序列和推导氨基酸序列,综合分析结构蛋白VP1的亲水性、可塑性、抗原指数以及表面可能性等参数,预测其潜在B细胞抗原表位并人工合成表位肽段,利用间接ELISA对潜在表位肽段进行筛选鉴定,结果显示,表位VP1a和VP1d为病毒株AF72结构蛋白VP1的优势B细胞表位,该结果为进一步的FMDV多表位疫苗研究提供有价值的参考依据.  相似文献   

2种制备三聚氰胺酶标抗原方法的比较     

王黎丽  余为一 《安徽农业科学》2011,39(22):13273-13274

[目的]对直接和间接法制备酶标记三聚氰胺抗原进行比较。[方法]2种方法均用辣根过氧化物酶和过碘酸钠法,但直接法是酶与三聚氰胺偶联,而间接法是用酶标记三聚氰胺与载体蛋白OVA的结合物。用上述方法分别制备了2种酶标抗原后,从酶活性、抗原性和效价方面对这2种酶标抗原做质量对比试验。[结果]间接法制备的HRP-OVA-MEL有抗原性,且效价达到1∶2 560以上,而直接法制备的HRP-MEL却没有抗原性,且效价较低。[结论]间接法制备的三聚氰胺酶标抗原的活性比直接法好。  相似文献   

不同土壤有机碳测定方法的比较   总被引：1，自引：1，他引：0  

唐伟祥  孟凡乔  张煜  赵自超 《土壤》2018,50(3):552-557

对土壤碳的准确定量是研究土壤肥力和农业固碳的前提。本文选择我国不同有机碳和无机碳含量的8个代表性土壤/沉积物样品,比较湿氧化法、酸洗法和间接法三类方法对有机碳含量的测定效果。研究结果表明:对有机碳含量为3.4~11.5 g/kg的土壤和沉积物样品,湿氧化法有较低的平均相对误差(0.32%~4.72%)和变异系数(0.40%~1.22%),间接法的平均相对误差(5.16%~20.46%)和变异系数(0.60%~7.00%)都较高,而酸洗法介于这两个方法之间。说明在保持稳定准确的加热温度情况下,湿氧化法对土壤/沉积物有机碳含量的测定比较稳定和准确,且操作简单。对大部分样品,由于酸对土壤无机碳难以充分溶解,造成酸洗法和间接法的有机碳测定值偏高,且变异系数较大。  相似文献   

食品总铬质量浓度icELISA试剂盒的研制及初步应用     

王亚楠  王晓斐  张雅凌  张海棠  王自良 《西北农业学报》2017,(12):1868-1876

为研制食品中总铬质量浓度检测间接竞争ELISA(icELISA)试剂盒(Cr-Kit)。利用蛋白质连接技术合成免疫原Cr~(3+)-iEDTA-BSA,紫外扫描(UV)和电感耦合等离子体原子发射光谱(ICP-AES)进行鉴定;用Cr~(3+)-iEDTA-BSA免疫BALB/c小鼠,细胞融合技术筛选抗Cr~(3+)-EDTA mAb细胞株,体内诱生腹水法制备Cr~(3+)-EDTA mAb,并对其特性进行分析;应用Cr~(3+)-EDTA mAb研制Cr-Kit,测定其性能并进行初步应用。结果表明,免疫原制备成功,Cr~(3+)-iEDTA-BSA中Cr~(3+)和BSA的质量浓度分别为140.8mg/L和5.81g/L;筛选出2A3C11、2A3D9、2A11G5共3株杂交瘤细胞株,其中2A11G5最好,经6次传代分泌抗体稳定,亲和常数(Ka)为2.69×109 L/mol,与其他重金属离子无交叉反应;Cr-Kit标准曲线的线性范围为1.0~264μg/L,检测限为1.0μg/L,IC50为8.37μg/L,河水样、大米样、面粉样和猪肉样的平均加标回收率分别为101.7%、96.47%、95.8%和84.3%,Cr-Kit与国标(GB5009.123-2014)石墨炉原子吸收光谱法(GFAAS)的符合率为100%。本研究成功研制了Cr-Kit,可满足食品总铬残留的检测要求。  相似文献   

"十二五"期间凤城市5种主要林业有害生物灾害损失评估     

杨庆寅 《安徽农业科学》2017,45(20)

根据凤城市"十二五"期间林业有害生物发生情况,筛选出美国白蛾、日本蚣干蚧、栗山天牛、杨干象、杨树烂皮病5种主要林业有害生物作为灾害损失评估对象,进行直接和间接经济损失评估计算。结果表明,凤城地区"十二五"期间5种主要林业有害生物造成的年均损失为61 376.777万元,其中,直接经济损失87.218万元,间接经济损失为61 289.559万元。  相似文献