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大肠杆菌O157及其Stx2免疫胶体金层析检测方法的建立 总被引:1,自引:0,他引:1
建立了大肠杆菌O157和志贺毒素2(Stx2)的双抗体夹心免疫胶体金层析法,并运用该方法对上海市兽医生物技术重点实验室保存的已鉴定的58株大肠杆菌进行了检测,评估了其特异性、敏感性和重复性.结果显示,大肠杆菌O157层析法的敏感性为1×105CFU/mL,特异性为87.5%,批间和批内变异系数分别为8.6%和0;Stx2层析法的检测下限为200μL细菌培养液,特异性为50.0%,批间和批内变异系数分别为50.0%和33.0%.表明,该免疫胶体金层析法能快速、简便、特异地检测大肠杆菌O157;结合对Stx2的检测,能快速检出产Stx2的大肠杆菌O157菌株. 相似文献
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在全国流行病学调查的基础上 ,选择四川等地流行广泛的鸭疫里氏杆菌 (RA) 1、2、4和5血清型菌株制备铝胶佐剂四价灭活疫苗。对疫苗经无菌检验及安全检验合格后 ,给 3日龄樱桃谷商品鸭每只颈背皮下注射 0 .5mL ,第 10、13和 16d免疫鸭对同源RA的攻击分别表现出 5 %~2 0 %、75 %~ 90 %和 10 0 %免疫保护力 ,第 2 3~ 30d免疫保护力开始下降。用间接ELISA对免疫鸭进行抗体消长规律检测 ,表明 3日龄樱桃谷鸭首免后第 2 3d抗体达到高峰 ,至 93日龄时仍能检出抗体 ;10日龄时进行二免后可产生更高的抗体水平 ,到第 65d时仍能保持较高抗体水平。疫苗免疫雏鸭能够显著 (P <0 .0 5 )或极显著 (P <0 .0 1)地增强其T细胞的免疫力 ,时间达 2周。经攻毒保护试验、抗体消长规律测定、T细胞免疫水平测定和田间试验 ,证明研制的鸭疫里氏杆菌病(RAI)四价灭活疫苗具有良好的安全性和免疫原性 ,能有效预防RAI的发生 相似文献
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《中国兽医科学》2020,(11)
为分离、鉴定猪盖他病毒(GETV)并对其灭活疫苗的免疫原性和免疫保护性进行评价,本研究对猪腹泻病样品进行了二代测序,用Vero细胞分离病毒并进行了qRT-PCR和IFA鉴定及遗传进化分析;用该分离株制备油乳剂灭活疫苗,并在猪体进行了免疫抗体水平测定和免疫攻毒保护研究。结果显示,二代测序检测出腹泻样品中存在GETV;经细胞分离、qRT-PCR和IFA鉴定证实,成功分离到一株猪源GETV,将其命名为GETV JS18株;该分离株与我国其他猪源分离株同属于基因Ⅲ型,与我国2017年分离株AH9192株的亲缘性最近,序列同源性为99.7%。GETV JS18株以0.005 MOI接种Vero细胞48 h毒价最高,达7.6±0.2 Log_(10)TCID_(50)/m L;该毒株制备的油乳剂灭活疫苗免疫仔猪,可有效阻止GETV感染引起的临床症状和平均日增重(ADWG)下降,降低GETV感染引起的病毒血症并阻止排毒和组织带毒,对攻毒保护率为100%;疫苗免疫后的平均中和抗体滴度最高可达9.8±0.8 Log2,且持续较高水平至少7个月。上述结果表明,本研究成功分离了一株猪源基因Ⅲ型GETV JS18株,其油佐剂灭活疫苗具有良好的免疫保护效力和免疫持续期。 相似文献
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为建立监测Cfr蛋白的方法奠定基础,采用PCR方法扩增cfr基因,并克隆入pEasy-T载体;测序确认后克隆入pET-28a(+)载体,构建重组质粒pET-28a-cfr,然后转化入大肠杆菌BL21(DE3)中并用IPTG诱导表达Cfr蛋白;用纯化的Cfr蛋白免疫BALB/c小鼠制备多克隆抗体,并对抗体进行鉴定。结果显示,成功克隆了全长cfr基因,构建了重组质粒pET-28a-cfr,并诱导了Cfr蛋白表达以及制备了小鼠抗Cfr蛋白的多克隆抗体;Western-blot法鉴定该抗体可特异性识别重组蛋白Cfr;ELISA法测定抗体效价为1∶64 000。结果表明,用大肠杆菌BL21(DE3)能够成功表达Cfr蛋白,并且获得了高特异性、高效价的小鼠抗Cfr蛋白的多克隆抗体。 相似文献
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《中国兽医科学》2020,(11)
为建立奶牛血清中油酸(OA)含量的免疫学检测方法,将油酸用活泼酯法分别与牛血清白蛋白(BSA)和卵清白蛋白(OVA)进行偶联,制备人工抗原和包被原;利用薄层色谱法(TLC)和红外光谱法鉴定人工抗原合成是否成功;用OA-BSA常规免疫6周龄雌性BALB/c小鼠,取小鼠脾细胞和骨髓瘤细胞进行融合,用间接ELISA筛选出阳性细胞株,再用有限稀释法进行亚克隆;采用小鼠体内诱生法制备腹水,Hi Trap Protein G HP纯化柱纯化腹水,并对纯化后的腹水用ELISA检测抗体效价,以及抗体亚型鉴定。结果表明,TLC监测抗原合成过程中有新物质生成,红外图谱显示OA-BSA波谱形态与BSA相似且拥有OA的特征峰;小鼠血清抗体效价为1∶12 800,用ELISA方法筛选出1株细胞G-1,抗体亚型为Ig G1型。为建立油酸的免疫学检测方法奠定了基础。 相似文献
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Zhang Jian 《现代国际关系(英文版)》2008,18(1):74-87
The EU's strategy for Russia since the end of the cold war has to a large extent failed, and the direct consequences are the currently deadlocked EU-Russia relationship. There are three reasons for this: the EU and Russia's respective strength and status relative to each other have been changing since the beginning of this century; the EU's strategy towards Russia has been more fragmenting in recently years; and the American factor. The EU-Russia relations could continue to be in poor shape in the years to come, and it is very likely sometimes even further worsening. But the two sides would refrain from extreme measures and prevent their relations from sliding into real cold war. 相似文献
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Zhang Zhixin 《现代国际关系(英文版)》2008,18(1):44-61
In this paper, the author has made a thorough study over the new generation of American China Watchers. Most of the pages are devoted to a detailed comparison between the old generation and the young generation of American China Watchers regarding to their background, education, and views over China. At the end of the paper, the author also gives some policy suggestions for both the Chinese and U.S. government based on the findings in this study. 相似文献
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Zhong Shujia 《现代国际关系(英文版)》2008,18(1):62-73
1. Introduction
On the morning of April 1st, 2001, a U.S. EP-3 surveillance aircraft was conducting an intelligence flight along the Chinese territorial border in the airspace south-east of China's Hainan Island. Two Chinese fighter jets intercepted the spy plane and followed its flight, as is considered ordinary international practice. For reasons unknown, the EP-3 airplane made a sudden maneuver and crashed into one of the Chinese planes. The Chinese plane dropped into the sea, the pilot missing. The U.S. spy plane was also severely damaged. It intruded into Chinese territorial space and landed on Hainan Island without official permission from the Chinese side. After landing, the spy plane and 24 crew on board were detained by the Chinese authorities. This air collision episode was the first of its kind in the China-U.S. relations and an urgent diplomatic crisis ensued. 相似文献
On the morning of April 1st, 2001, a U.S. EP-3 surveillance aircraft was conducting an intelligence flight along the Chinese territorial border in the airspace south-east of China's Hainan Island. Two Chinese fighter jets intercepted the spy plane and followed its flight, as is considered ordinary international practice. For reasons unknown, the EP-3 airplane made a sudden maneuver and crashed into one of the Chinese planes. The Chinese plane dropped into the sea, the pilot missing. The U.S. spy plane was also severely damaged. It intruded into Chinese territorial space and landed on Hainan Island without official permission from the Chinese side. After landing, the spy plane and 24 crew on board were detained by the Chinese authorities. This air collision episode was the first of its kind in the China-U.S. relations and an urgent diplomatic crisis ensued. 相似文献
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Ruan Zongze 《中国国际问题研究》2014,(2)
正The new central leadership has highlighted the strategic importance of better relations with neighboring countries and the construction of a solid foundation for China’s neighborhood strategy so as to create favorable conditions for Chinese development.1China is strengthening top-level strategy in foreign relations,focusing on global-level relations and staying committed to relations 相似文献
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正Ruicheng Group was founded in Qiqihar,a city located in one of the three major ‘Chernozem belts’ in the world,the fertile Songnen Plain,and one known as the ‘home of red-crowned cranes’ Over the years,our business developed from being only one supermarket in the beginning to a corporate group operating in multiple sectors,from supermarket chain and real estate to property management and preschool education. 相似文献
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江时学 《中国国际问题研究》2014,(4)
China and Brazil established a strategic partnership in 1993. The bilateral ties have seen rapid development in almost all areas. However, there are eight myths surrounding the China-Brazil relations. These myths are the result of a lack of mutual understanding as well as the misconceptions result misconceptions of the western observers and media. These misunderstandings have of have jeopardized images of the China-Brazil relations and also hindered further development of their bilateral relationship. 相似文献
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陈东晓 《中国国际问题研究》2014,(5)
The successful hosting of the CICA summit has drawn great attention The attention to China's "host diplomacy". Analyzing new features of China's "host to "host diplomacy" during this new period, this paper will look at the opportunities and challenges facing China's "host diplomacy" and discuss the problems that China should address in order to do a better job in this regard and bring its "host diplomatic advantages" into full play. 相似文献
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高祖贵 《中国国际问题研究》2014,(3)
Since the beginning of the 21st century, the overall rise of the Asian geographic plate has been particularly prominently. The overall rise of Asia reflects the improvement of a large number of developing countries in Asia. The long-occupied aggressive status of the United States and Europe is weakening, and the overall rise of Asia appears more highlighted, and its impact upon the world seems more intense. 相似文献
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正On the morning of April 2,2014,CAFIU held the 11th Executive Council Meeting in Beijing.Present at the Meeting were Member of the Politburo of the 17th CPC Central Committee and Vice-Chairman of the 11th CPPCC National Committee Mr.Wang Gang,ViceChairman of the 11th NPC Standing Committee and President of CAFIU Mr.Zhou Tienong,ViceChairpersons of the NPC Standing Committee Mme 相似文献
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正I am very pleased to attend the 11th Executive Council Meeting of CAFIU,and would like to express my hearty congratulations to President Yan Junqi,the new leadership and the newly-elected Executive Council.I also want to express my sincere thanks for choosing me to be the Honorary President of CAFIU! 相似文献