猪细小病毒、猪圆环病毒2型和伪狂犬病病毒多重PCR检测方法的建立

根据GenBank中登录的猪细小病毒(PPV)、猪圆环病毒2型(PCV2)和伪狂犬病病毒(PRV)核苷酸序列,分别设计并合成了3对特异性扩增PPV、PCV2和PRV的引物,扩增片段长度分别为531、466和277bp。经过优化条件,建立了检测PPV、PCV2和PRV的多重PCR方法。采用该PCR方法对23份自然感染病猪样品进行检测,结果表明,该多重PCR检测结果与单一PCR检测结果的符合率为100%,同时对临床样品中PPV、PCV2和PRV阳性PCR产物进行测序分析,发现PPV、PCV2和PRV的PCR产物序列与GenBank中公布序列的同源性均达98%以上,证实该多重PCR检测的阳性结果为上述3种病毒。表明,建立的多重PCR方法可用于这3种病毒的临床诊断和流行病学调查。     

五种重要猪病病毒基因芯片探针的建立及其应用

将猪繁殖与呼吸综合征病毒(PRRSV)、猪瘟病毒(CSFV)、猪细小病毒(PPV)、猪伪狂犬病痛毒(PRV)、猪圆环病毒2型(PCV-2)的特异cDNA片段作为探针点制于氨基修饰的玻片上,以这5种细胞毒的核酸作为模板,进行不对称PCR扩增,制备靶物,经绿色荧光染料Cy3标记后,分别与cDNA微阵列的芯片探针进行杂交,杂交信号经Genepix 4000A扫描仪扫描.结果显示,该芯片探针可以特异地与Cy3标记的这5种靶物结合.用该基因芯片对100份现地采集的猪全血样品进行检测,检出PRRSV阳性样品26份,PCV-2阳性样品47份,PRRSV和PCV-2混合感染阳性样品17份,PPV阳性样品5份,PRV阳性样品2份,CSFV阳性样品20份.表明,本试验研制的cDNA芯片探针可以特异地检测这5种猪病病毒,具有良好的诊断应用前景.     

三种猪繁殖障碍性病毒混合感染的分子生物学调查

根据GenBank登录的PRRSVORF7、PCV 2ORF1基因及PRVTK基因的核苷酸序列设计合成引物 ,建立了分别用于检测PRRSV、PRV及PCV 2的RT PCR及PCR方法。应用该方法对 77份可疑病料进行了检测 ,以调查猪群中PCV 2与PRRSV和 (或 )PRV混合感染情况。结果表明 ,2 4份样品表现为PRRSV与PCV 2混合感染 ,占样品总数的 31.2 % ;17份样品检测为PRRSV与PRV混合感染 ;9份样品表现为PCV 2与PRV混合感染 ,占 11.7% ;另外 ,还有一定比例的三重感染 ,共 8个样品 ,占 10 .4 %。结果显示 ,猪群中PRRSV、PRV与PCV 2的混合感染现象比较普遍     

长江中下游地区规模化猪场猪繁殖障碍性疾病的病原学检测

应用PCR与RT-PCR方法,检测了127份来自于长江中下游地区7省市24个规模化猪场患繁殖障碍的病猪的病料,检测病原包括猪生殖与呼吸综合征病毒(PRRSV)、伪狂犬病病毒(PRV)、猪瘟病毒(CSFV)、猪圆环病毒2型(PCV-2)、猪细小病毒(PPV)及日本脑炎病毒(JEV).结果显示,在上述127份样品中,有10份样品为PRRSV单独感染,占检测样品数的7.87%,有54份样品为PRRSV与PPV混合感染,占样品总数的42.52%,有50份样品为PRRSV与PCV-2混合感染,占样品总数的39.37%;有15份样品为PRRSV与PRV混合感染,占样品总数的11.81%;有8份样品为PRRSV与JEV混合感染,阳性率为6.30%;有18份样品为PRRSV与CSFV混合感染,占样品总数的14.17%.提示,长江中下游地区规模化猪场的猪群普遍存在PRRSV与PPV、PCV-2、JEV及PRV的混合感染.     

猪圆环病毒2型和猪生殖与呼吸综合征病毒混合感染的检测

对采自甘肃省天水市某猪场的24份病猪组织器官样品(肺、脾、肾和淋巴结)进行了病毒分离,同时设计特异性引物,建立了分别检测猪圆环病毒2型(PCV-2)和猪生殖与呼吸综合征病毒(PRRSV)的PCR和RT-PCR方法,应用ELISA方法检测了血清中PCV-2及PRRSV抗体,以调查该猪场PCV-2和PRRSV混合感染的情况.结果显示,有14份样品均扩增出了PCV一2和PRRSV的目的基因片段,且PCV-2及PRRSV抗体均呈阳性.证实,该猪场出现了PCV-2和PRRSV的混合感染,阳性率为58.33%.     

猪圆环病毒2型猪伪狂犬病病毒和猪细小病毒三重PCR检测方法的建立

针对猪圆环病毒2型(PCV2)ORF2、猪伪狂犬病病毒(PRV)gD和猪细小病毒(PPV)VP2基因保守区域,设计了特异性引物,在分别建立单一病毒基因PCR检测方法的基础上,通过对3组引物的比例和浓度、dNTPs和Mg2 浓度、退火温度等条件的优化,建立了针对上述3种病毒的三重PCR检测方法。敏感性试验表明,应用该方法最低可检测到4×10-4ng/μL的PRV双链DNA模板和2×10-5ng/μL的PCV2和PPV单链DNA模板。对21份自然感染病猪样品的检测结果表明,该三重PCR检测结果与单一PCR检测的结果完全符合。试验结果显示,建立的三重PCR方法可用于3种DNA病毒的同时检测,具有快速、准确的特点,有临床应用前景。     

PCV2-PRV-PRRSV-CSFV多重PCR检测方法的建立

根据GenBank中已发表的猪圆环病毒2型(PCV2)、伪狂犬病病毒(PRV)、猪繁殖与呼吸综合征病毒(PRRSV)和猪瘟病毒(CSFV)的有关基因序列,设计4对引物分别用于扩增PCV2的ORF2基因、PRV的gH基因、PRRSV的ORF7基因和CSFV的5′UTR基因的目的片段,建立了同时检测4种病毒的多重PCR。敏感性和特异性试验结果表明,该方法对这4种病毒的最低核酸检出量分别为48.1pg(PCV2)、29.6pg(PRV)、54.2pg(PRRSV)和62.8pg(CSFV)。该方法可同时扩增440bp(PCV2)、359bp(PRV)、321bp(PRRSV)和186bp(CSFV)的特异性片段,而对牛流行性腹泻病毒、猪流行性腹泻病毒和猪细小病毒的检测结果均为阴性。多重PCR与单一PCR的符合率为100%。结果表明,该方法可用于这4种病毒的临床快速检测与流行病学调查。     

四种猪呼吸道疾病综合征病原多重PCR检测方法的建立及应用

为建立同时检测猪繁殖与呼吸综合征病毒(PRRSV)、猪圆环病毒2型(PCV2)、胸膜肺炎放线杆菌(APP)和猪肺炎支原体(M.hyo)的多重PCR方法,分别针对PRRSV的ORF1a基因、PCV2的ORF2基因、APP的OMLA基因和M.hyo的P46基因的保守区域设计4对特异性引物,优化反应体系及条件,建立了可同时扩增PRRSV(225bp)、PCV2(337bp)、APP(107bp)、M.hyo(176bp)相应基因的四重PCR方法。结果显示,该方法能单管同时特异地鉴别检测PRRSV、PCV2、APP、M.hyo 4种病原体,对猪瘟病毒、副猪嗜血杆菌、猪呼吸道冠状病毒、猪流感病毒、猪细环病毒及猪细小病毒等无特异性扩增,说明该PCR具有良好的特异性;方法的灵敏度分别为195、272、241、321pg/μL。应用该方法对重庆市的61份样品进行检测,其中PRRSV、PCV2、APP、M.hyo的阳性率分别为22.9%(14/61)、18.0%(11/61)、8.2%(5/61)和18.0%(11/61)。该方法的建立为临床上4个病原的快速鉴别诊断提供了有力的技术手段。     

广西猪圆环病毒2型感染的流行病学调查

结合流行病学、临床症状、病理变化,采用PCR技术,对2004年1月至2005年5月采自广西14个市97个疑似猪圆环病毒2型(PCV2)感染发病猪场的197份组织病料(脾、肺、淋巴结)进行了PCV2检测;同时,对鉴定为PCV2阳性的组织病料和猪场进行了猪生殖与呼吸综合征病毒(PRRSV)、猪瘟病毒(CSFV)、猪流感病毒(SIV)和猪伪狂犬病病毒(PRV)的检测,另外,对6个地(市)11个生猪屠宰场采集的外观健康屠宰猪的295份组织样品(脾、肺、淋巴结)进行了PCV2检测。结果显示,在197份组织样品中检出PCV2阳性病料108份,平均阳性率为54.82%(108/197),阳性猪场62个,平均阳性率为63.92%(62/97)。PCV2与PRRSV、CSFV、SIV、PRV混合感染的组织病料总阳性率为42.13%(83/197),混合感染的猪场总阳性率为57.73%(56/97)。从21头外观健康屠宰猪的组织样品中检测到PCV2,阳性率为7.10%。由此可见,PCV2感染在广西猪群中已普遍存在,混合感染和健康带毒现象使病情更加复杂。     

用多重PCR鉴别猪伪狂犬病野毒与疫苗毒的研究

根据基因库中的猪伪狂犬病病毒(PRV)各基因的序列,设计了与PRV的 gB、gD、gE基因序列互补的3对引物。对样品中的PRV DNA模板进行了多重PCR扩增及反应条件的优化,结果同时得到与设计相符合的3条特异性条带,分别为549 bp(gB)、429 bp(gD)、366 bp(gE)。用这3对引物对三基因缺失疫苗毒的样品DNA模板进行多次多重 PCR扩增,均能稳定得到与设计相符合的2条特异性条带。敏感性试验结果表明,多重 PCR可以检测到 106 pg三基因缺失疫苗毒或756 pg PRV野毒的核酸模板量。特异性试验结果表明,以正常对照细胞及猪圆环病毒和猪细小病毒DNA为模板进行多重PCR扩增,均无任何条带。     

The roles of,and relationships between,expatriates, volunteers,and local development workers

This article reports on exploratory research based on interviews with expatriate and local aid workers employed by local and international NGOs in Phnom Penh, Cambodia. Thematic analysis of the interviews found that personnel were placed in groups based on their job category – consultant, volunteer, or permanent staff – regardless of experience. These categories logically reflect each worker's pay level, purpose, and role, but they may also have an implicit power meaning which reinforces group differences and inhibits inter-group relationships. Relationship building was reported to be the most important factor contributing to the success of capacity-development initiatives. Four sub-themes were identified: communication, friendship, reciprocal learning/teaching, and confidence.     

Britain, France, and America's Year of Europe, 1973

Britain's entry into the European Community in 1973 coincided with an American initiative aimed at redefining relations between the United States and Western Europe. This confronted British diplomats with a serious dilemma. They wished to maintain close collaboration with Washington and, for the sake of European unity, to expand on their recently achieved reconciliation with France, a country whose Gaullist elite rejected any further institutionalization of transatlantic relations. French reluctance to engage in a constructive dialogue with the Americans resulted in a fractious debate over the drafting of two seemingly innocuous declarations, and this was exacerbated by the mixed response of the Europeans to the outbreak of the fourth Arab-Israeli war and the ensuing energy crisis. Finally, at the Washington energy conference of February 1974, the British chose to work with the Americans, rather than the French, in seeking to mitigate the economic impact of OPEC's oil policies.     

Britain,France, and America's Year of Europe, 1973

Britain's entry into the European Community in 1973 coincided with an American initiative aimed at redefining relations between the United States and Western Europe. This confronted British diplomats with a serious dilemma. They wished to maintain close collaboration with Washington and, for the sake of European unity, to expand on their recently achieved reconciliation with France, a country whose Gaullist elite rejected any further institutionalization of transatlantic relations. French reluctance to engage in a constructive dialogue with the Americans resulted in a fractious debate over the drafting of two seemingly innocuous declarations, and this was exacerbated by the mixed response of the Europeans to the outbreak of the fourth Arab–Israeli war and the ensuing energy crisis. Finally, at the Washington energy conference of February 1974, the British chose to work with the Americans, rather than the French, in seeking to mitigate the economic impact of OPEC's oil policies.     

Terrorism,counter-terrorism,and critique: opportunities,examples, and implications

This article explores the parameters, value and limitations of different critical strategies for those dissatisfied with the contemporary politics of terror. It argues, first, that the prominent (counter-)terrorism paradigm – in which terrorism is approached as a ubiquitous and very specific security challenge meriting appropriately exceptional responses – is far more critiqued than we might anticipate. And, second, that such critiques – which can be found across political language, popular culture, everyday life, and beyond – employ distinct critical resources to serve varying ends. Reflecting on these, the article offers a new heuristic distinguishing five critical strategies which seek to: (i) repudiate; (ii) question; (iii) subvert; (iv) replace, or (v) deconstruct the prominent counter-terrorism paradigm. This typology, it argues, offers scope for optimism and strategic resources for those attracted to a critical terrorism studies project going forward.     

Population, Prophets, Pensions, and Politics

As immigrant populations in Western welfare states grow at a faster rate than the native populations, whose birthrates have declined dramatically in recent years, Europe's tradition of democracy and tolerance is threatened. The reasons for the birthrate decline and the resultant aging of the native population are many and complex, but one important contributing factor is the pay-as-you-go pension system, which reduces people's immediate dependence on children. Moreover, the payroll and social security taxes that support the welfare state reduce the earnings capacity of men and women of the traditional age for having children, thus pushing down the birthrate. It is time to consider whether modern Europe's small-size families and high pensions are sustainable     

“印太战略”:以美印日澳的战略逻辑、利益与策略选择为分析视角

"印太战略"是美国均势+有限遏制+规制+话语诋毁的混合型战略。它意在前沿、中间地带与后方的三层构建不对称的对华遏制性力量,突破中美双边、单区域战略竞争的狭窄战略空间;在多地域与多维度孤立中国,拉拢更多国家在产业价值链、数字经济等层面与中国切割;制订基础建设投资透明化的新标准,嵌入有利于美国利益的规制;通过国际法、舆论等软性手段抹黑中国的"一带一路"与海权发展。虽然共同逻辑都是制衡中国崛起,但日澳印在策略上强调非对抗性。其中避免成为中国优先反制是日本重要的策略目标;印度在拉达克边界冲突后尚没有形成一个完全倒向美国的战略,因为这必定使其背上沉重的战略包袱,成为美国的附庸;澳大利亚作为地缘政治影响有限的中等国家,在美国反华行动中冲在最前列已使其利益严重受损。而"印太战略"与东盟所推动的包容性、合作安全理念也有着本质的区别,东盟要在"印太地区"发挥中心领导作用,定位"印太"为对话与合作区域。所以由于东盟的抵制、印日澳的消极政策,这个战略形成的攻势将很难保持可持续性,其长期前景并不看好。     

Indigeneity,Ethnicity, and the State: Australia,Fiji, and New Zealand

This article draws on the politics of indigeneity to distinguish the claims of first occupancy from simple ethnic identity politics, illustrating that relative political marginalization in Australasia is not so much a function of minority status but of indigeneity itself. The politics of indigeneity's aim is to create political space for self-determination and a particular indigenous share in the sovereign authority of the nation-state itself. The Australasian states are compared with Fiji to demonstrate that the significance of historical constraints on political authority transcend the withdrawal of a colonial power and the restoration of collective indigenous majority population status.     

Demographics, Religion, and the Future of Europe

Immigration and changing demographic trends mean that Europe will in the very near future inevitably be transformed, culturally and politically. As in the Cold War, it again represents a critical theater for rivalry, but this time it is between Christianity, Islam, and secularism. European nations will either be the sites of religious conflict and violence that sets Muslim minorities against secular states and Muslim communities against Christian neighbors, or it could become the birthplace of a liberalized and modernized Islam that could in turn transform the religion worldwide. We urgently need to understand the developing contours of European religious beliefs and practices, and not just as they apply to Muslims, for the outcome of the rivalry there will have profound implications for the United States.