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Development of three real-time PCR assays to detect Bacillus anthracis and assessment of diagnostic utility. 炭疽芽孢杆菌三种实时PCR检测方法的建立及诊断效果评价。
Pub Date : 2013-01-23 DOI: 10.4172/2157-2526.S3-009
Tanya M. Parsons, V. Cox, Angela E. Essex-Lopresti, M. G. Hartley, R. Lukaszewski, P. A. Rachwal, H. Stapleton, S. Weller
Three real-time PCR assays to detect Bacillus anthracis genetic targets (pXO1; pXO2 and chromosome) were developed. Two of the PCR assays (pXO1-MGB and Ba chr-MGB) were tested against DNA extracts produced from whole blood samples obtained from a replicated B. anthracis murine infection model. Across all three models 45 samples were tested in total, within which a subset of 41 samples were shown to contain B. anthracis by either PCR or microbiological culture. Using microbiological culture as an analogue of conventional blood culture (as used in clinical settings) the detection rates of PCR and blood culture were compared. In two of the murine models blood culture had a significantly higher detection rate than PCR (BA1, p=0.004; BA3, p=0.013). In the BA2 model there was no significant difference between the detection rates of PCR and blood culture.
三种实时PCR检测炭疽芽孢杆菌基因靶点(pXO1;pXO2和染色体)发育。两种PCR检测方法(pXO1-MGB和Ba hr- mgb)与从复制炭疽芽孢杆菌感染小鼠模型的全血样本中提取的DNA提取物进行了测试。在所有三种模型中,总共测试了45个样本,其中41个样本的一个子集通过PCR或微生物培养显示含有炭疽芽孢杆菌。将微生物培养作为常规血培养的模拟物(用于临床环境),比较PCR和血培养的检出率。在两种小鼠模型中,血培养的检出率显著高于PCR (BA1, p=0.004;BA3, p = 0.013)。在BA2模型中,PCR与血培养的检出率无显著差异。
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引用次数: 10
Bacteriophages against biothreat bacteria: diagnostic, environmental and therapeutic applications. 抗生物威胁细菌的噬菌体:诊断、环境和治疗应用。
Pub Date : 2013-01-21 DOI: 10.4172/2157-2526.S3-010
A. Filippov, Kirill V. Sergueev, M. Nikolich
Plague, anthrax and brucellosis are severe bacterial infections presenting a serious threat to public health. Their causative agents can be weaponized and a number of drug-resistant strains have been described. This requires improvement of existing and development of new methods of diagnostics, strain characterization, prophylaxis and therapy of these infections. This review article focuses on lytic bacteriophages (phages) active against Yersinia pestis, Bacillus anthracis and Brucella including the uses of phages for diagnostics, strain typing, specific decontamination, and antibacterial therapy.
鼠疫、炭疽和布鲁氏菌病是严重的细菌感染,对公众健康构成严重威胁。它们的病原体可以被武器化,并且已经描述了许多耐药菌株。这需要改进现有的和开发新的诊断方法、菌株特征、预防和治疗这些感染的方法。本文综述了对鼠疫耶尔森氏菌、炭疽芽孢杆菌和布鲁氏菌具有活性的溶菌体(噬菌体),包括噬菌体在诊断、菌株分型、特异性去污和抗菌治疗中的应用。
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引用次数: 13
Circulation of Multidrug Resistant Pathogens between Developed and Developing Countries: A New Frontier of Biodefense. Implication for Policy Makers 多药耐药病原体在发达国家和发展中国家之间的传播:生物防御的新前沿。对政策制定者的启示
Pub Date : 2013-01-01 DOI: 10.4172/2157-2526.1000E109
G. Ceccarelli, C. Ceccarelli, L. Pacifici
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引用次数: 4
Immunization with DNA Vaccine Expressing Gn Coupled to C3d Prevents Clinical Symptoms of Infection and Protects Mice against an Aerosol Rift Valley Fever Virus Infection. 表达Gn与C3d偶联的DNA疫苗免疫可预防临床感染症状并保护小鼠免受气溶胶裂谷热病毒感染
Pub Date : 2013-01-01 DOI: 10.4172/2157-2526.S3-006
N. Bhardwaj, Brooke R. Pierce, T. Ross
Rift Valley fever virus (RVFV) is the causative agent of Rift Valley fever (RVF) and is an emerging infectious disease of zoonotic potential. However, aerosolization of RVFV has been proposed as a potential bioweapon and most vaccines have not been tested against aerosolized RVFV challenge. Previously, two vaccine platforms (DNA plasmids and alphavirus replicons) expressing a soluble form of the RVFV Gn glycoprotein alone or fused to three copies of complement protein, C3d, protected mice against an intraperitoneal (IP) RVFV infection. In this study, both vaccine candidates were used to determine the protective efficacy against an aerosolized RVFV challenge. Each vaccine was administered to mice alone or in a heterologous prime/replicon boost strategy and anti-RVFV immune responses were assessed. DNA plasmids expressing Gn-C3d and alphavirus replicons expressing Gn elicited high titer neutralizing antibodies that were similar to titers elicited by the live-attenuated MP12 virus. However, only Gn-C3d- DNA vaccine completely protected mice against virulent aerosolized RVFV challenge. Most mice receiving replicon based vaccines succumbed to RVFV infection. Surprisingly, even though live-attenuated MP12 vaccine protected mice against IP challenge, MP12 did not provide complete protection against aerosolized RVFV infection. Therefore, vaccine candidates that are effective against peripheral challenge should be tested against aerosolized challenge to determine the complete protection profile, since any bioterrorism attack using RVFV would most likely be in the form of an aerosol.
裂谷热病毒(RVFV)是裂谷热(RVF)的病原体,是一种具有人畜共患潜力的新发传染病。然而,裂谷热病毒的雾化已被认为是一种潜在的生物武器,大多数疫苗尚未针对雾化的裂谷热病毒挑战进行过测试。以前,两种疫苗平台(DNA质粒和甲病毒复制子)单独表达一种可溶性形式的RVFV Gn糖蛋白或与三拷贝补体蛋白C3d融合,可保护小鼠免受腹腔(IP) RVFV感染。在这项研究中,使用这两种候选疫苗来确定对雾化的裂谷热病毒攻击的保护功效。每种疫苗单独或以异源引物/复制子增强策略给予小鼠,并评估抗裂谷热病毒免疫反应。表达Gn- c3d的DNA质粒和表达Gn的甲病毒复制子引发了高滴度的中和抗体,其滴度与MP12减毒病毒引发的滴度相似。然而,只有Gn-C3d- DNA疫苗完全保护小鼠免受毒力雾化的RVFV攻击。大多数接受复制子疫苗的小鼠死于裂谷热病毒感染。令人惊讶的是,尽管MP12减毒活疫苗可以保护小鼠免受IP攻击,但MP12并不能完全保护小鼠免受雾化的RVFV感染。因此,应对外周病毒攻击有效的候选疫苗应进行雾化攻击测试,以确定完整的保护情况,因为任何使用裂谷热病毒的生物恐怖袭击都极有可能以气溶胶的形式进行。
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引用次数: 0
Evidence for the Source of the 2001 Attack Anthrax 2001年袭击炭疽病毒来源的证据
Pub Date : 2013-01-01 DOI: 10.4172/2157-2526.S3-008
M. Hugh-Jones, B. H. Rosenberg, Stuart Jacobsen
The elemental composition of the 2001 attack anthrax presents critical clues that were not considered or were misinterpreted throughout the original investigation. Extensive experimental data released by the FBI after the anthrax case was closed make it possible to trace some of the implications of these clues: the substantial presence of tin, a toxic material that must have been added subsequent to growth, and a uniquely high content of silicon in the attack spores. No Bacillus spore preparations other than the attack anthrax have ever been found to contain such a high level of silicon, although some surrogate spore powders prepared at Dugway following FBI instructions have been cited as evidence that high levels of silicon can be reproduced; however, examination of the experimental data reveals that the silicon in these samples was unquestionably an artifact. The elemental evidence suggests that the attack spores had been coated with silicone (a polysiloxane) in the presence of tin, which catalyzes the cross-linking of polysiloxane chains needed to form an encapsulating coating on the spore coat. Microencapsulation helps protect biological agents from damage during atmospheric exposure and from the body’s defenses during infection, and would defeat some detection methods. Microencapsulation, which would explain the location and amounts of both tin and silicon in the attack spores, requires special expertise and sophisticated facilities. DOD-sponsored projects explicitly involving microencapsulation at DARPA, Dugway and perhaps elsewhere were spelled out publicly in budget documents in 1999 and thereafter, and executed at the very time of the anthrax attacks. Both the Dugway laboratory and Battelle Memorial Institute, a sub-contractor at Dugway, had extensive experience in making Bacillus spore powders; both had access to Bacillus anthracis genetically matching the attack spores; both could have made the attack spores legally for institutions conducting biodefense activities that required microencapsulated spores. Furthermore, a small but significant amount of tin, about 4% of that in the attack spores, has been found in some surrogate spore products made at Dugway. A measureable tin content has not been found in any other Bacillus spores except the attack spores. The tin in the Dugway surrogates may have been a remnant, indicative of earlier, classified work. Avoidance of governmentsponsored, classified research may account for some of the limitations of the investigation.
2001年袭击炭疽的元素组成提供了在最初的调查中没有被考虑或被误解的关键线索。炭疽热案件结束后,联邦调查局公布了大量的实验数据,使得追踪这些线索的一些含义成为可能:锡的大量存在,一种有毒物质,一定是在生长后添加的,以及攻击孢子中独特的高含量硅。除了攻击炭疽菌外,从未发现芽孢杆菌孢子制剂含有如此高水平的硅,尽管在杜格威按照联邦调查局的指示制备的一些替代孢子粉末被引用为可以复制高水平硅的证据;然而,对实验数据的检查表明,这些样品中的硅毫无疑问是人造的。元素证据表明,攻击孢子在锡存在的情况下被硅酮(一种聚硅氧烷)包裹,锡催化聚硅氧烷链的交联,形成孢子外壳上的封装涂层。微胶囊化有助于保护生物制剂在暴露于大气中时不受破坏,在感染时不受人体防御的伤害,并且会击败一些检测方法。微胶囊化可以解释攻击孢子中锡和硅的位置和数量,这需要特殊的专业知识和复杂的设备。国防部资助的项目明确涉及DARPA、杜格威(Dugway)和其他地方的微胶囊技术,这些项目在1999年及之后的预算文件中被公开列出,并在炭疽病毒袭击发生的同一时间执行。杜格威实验室和杜格威的分包商巴特尔纪念研究所在制造芽孢杆菌孢子粉方面都有丰富的经验;他们都能接触到基因上与攻击孢子匹配的炭疽芽孢杆菌;这两家机构都可以合法地制造攻击孢子,用于进行需要微封装孢子的生物防御活动。此外,在杜格威制造的一些替代孢子产品中发现了少量但数量可观的锡,约占攻击孢子中锡的4%。除攻击孢子外,在其他芽孢杆菌孢子中未发现可测量的锡含量。杜格威替身里的锡可能是残留的,表明了早期的机密工作。避免政府资助的机密研究可能是调查的一些局限性的原因。
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引用次数: 4
A Need for Emphasis on Local Leadership in Emergency Management 需要强调地方在应急管理中的领导作用
Pub Date : 2012-12-03 DOI: 10.4172/2157-2526.S5-E001
G. Avery
ISSN:2157-2526 JBTBD, an open access journal J Bioterr Biodef Bioterrorism: Disaster preparedness education and public health In 2009, I participated in the Joint Urban Warrior exercise sponsored by the Marine Corps War fighting Laboratory and the Joint Forces Command-Joint Irregular Warfare Center, focusing on the problem of integrating military and civilian efforts in stability operations. In my analytical cell, the question arose as to where the necessary expertise could be found in civilian agencies, and in our discussions, it quickly became apparent that, for the most part, the applied skills were found not in federal civilian agencies, but in state and local government agencies. Besides representing a lesson lost within the military Civil Affairs community (in the Second World War Civil Affairs and Military Government soldiers were largely recruited from academia and sub-national government officials), but also an important cue that even in national security areas, the federal government may not always know best. A state extension officer, for example, may well know more about practical aspects of crop production than a federal USDA expert in agricultural price supports. In areas like biodefense or emergency preparedness, intricately entwined with public health, local officials are also likely to be the key players in effective incident response.
2009年,我参加了由海军陆战队作战实验室和联合部队司令部联合非正规作战中心主办的“联合城市勇士”演习,重点研究了在维稳作战中军民结合的问题。在我的分析小组中,出现了一个问题,即在民事机构中哪里可以找到必要的专业知识,在我们的讨论中,很快就发现,在大多数情况下,应用技能不在联邦民事机构中找到,而是在州和地方政府机构中找到。除了代表了军事民政界失去的教训(在第二次世界大战中,民政和军政府的士兵主要是从学术界和地方政府官员中招募的),而且也是一个重要的提示,即即使在国家安全领域,联邦政府也可能并不总是最了解的。例如,一个州的推广官员可能比联邦农业部在农业价格支持方面的专家更了解农作物生产的实际方面。在生物防御或应急准备等与公共卫生错综复杂地交织在一起的领域,地方官员也可能是有效应对事件的关键人物。
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引用次数: 1
Accelerated manufacturing of large-scale, full-length, human-like glycosylated antibodies for bio-defense. 加速大规模、全长、类人糖基化生物防御抗体的生产。
Pub Date : 2012-11-29 DOI: 10.4172/2157-2526.1000122
Christopher M. Warner, M. Croughan
From the point of DNA sequence confirmation until production of meaningful clinical quantities of novel therapeutics, current manufacturing systems for many glycosylated proteins require several months of development. Consequently, in the event of mass-casualty epidemics, current systems will fail to provide sufficient and timely quantities of emergency medical counter measures. As the identity of many new biological threats are unlikely to be known in advance, pre-emptive manufacturing and stockpiling of countermeasures cannot always be performed. Preparedness for all biological catastrophes requires a radical solution to replace the current slow scale-up and manufacture of certain lifesaving medical countermeasures. Current clinical and commercial manufacturing methods for glycosylated proteins use stable cell lines for protein expression, wherein the gene coding for the protein of interest is stably integrated into the host cell genome. The generation, identification, banking, testing, and scale-up of suitable stable clones generally takes many months. Because this development time is not compatible with emergency manufacturing, an alternative method for rapid production of medical counter measure antibodies is needed. One such potential technique is transient gene expression. Transient gene expression is a common approach for production of research-grade antibodies. It is frequently used to generate milligram to gram quantities of material within two to three weeks of DNA sequence confirmation. In the past, transient systems have been considered for emergency production of large quantities of antibodies, but dismissed due to low titers, high cost of DNA, uncertain regulatory environment, and the lack of sufficient, available manufacturing capacity. Recent developments, however, have substantially enhanced the viability of such an approach. This article will explore these developments and investigate the use of transient gene expression for rapid production of antibody-based medical countermeasures.
从DNA序列确认到生产有意义的临床数量的新疗法,目前许多糖基化蛋白的制造系统需要几个月的开发。因此,在发生大规模伤亡流行病时,现有系统将无法提供足够和及时的紧急医疗应对措施。由于许多新的生物威胁的性质不太可能事先知道,因此不可能总是先发制人地制造和储存对抗措施。为所有生物灾难做好准备需要一个彻底的解决办法,以取代目前缓慢扩大和制造某些挽救生命的医疗对策的做法。目前的临床和商业糖基化蛋白的制造方法使用稳定的细胞系进行蛋白表达,其中目标蛋白的基因编码稳定地整合到宿主细胞基因组中。生成、识别、存储、测试和扩展合适的稳定克隆通常需要几个月的时间。由于这种开发时间与应急生产不兼容,因此需要一种快速生产医疗对抗抗体的替代方法。其中一个潜在的技术是瞬时基因表达。瞬时基因表达是生产研究级抗体的常用方法。它通常用于在DNA序列确认后的两到三周内生成毫克到克的物质。过去,瞬态系统曾被考虑用于大量抗体的紧急生产,但由于低效价、DNA成本高、监管环境不确定以及缺乏足够的、可用的制造能力而被忽视。然而,最近的事态发展大大加强了这种办法的可行性。本文将探讨这些发展,并研究利用瞬时基因表达快速生产基于抗体的医疗对策。
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引用次数: 5
Allergic Examination for Biodefense 生物防御的过敏检查
Pub Date : 2012-11-27 DOI: 10.4172/2157-2526.1000E108
N. Oiso
Until now, various case reports and case series of allergic sensitization from novel ingredients in new industrial and consumer products have been reported. The scientific reports have been prompting the manufacturers to eliminate sensitizers provoking frequent allergic reaction and/or severe clinical symptom from products. The adequate cycle provides better situation for people living in the world. We need to keep in mind that more victims may be sensitized if decision to eliminate the hazardous components is delayed.
到目前为止,已经有各种各样的病例报告和病例系列报道了新的工业和消费产品中的新成分引起的过敏致敏。科学报告已经促使制造商从产品中去除引起频繁过敏反应和/或严重临床症状的致敏剂。充足的周期为生活在世界上的人们提供了更好的环境。我们需要记住,如果推迟消除危险成分的决定,可能会有更多的受害者受到影响。
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引用次数: 1
Lack of Adequate Surveillance of Biological Threats is a Peril to Global Public Health 缺乏对生物威胁的充分监测是对全球公共卫生的一种危害
Pub Date : 2012-11-24 DOI: 10.4172/2157-2526.S4-E001
Mingtao Zeng, Maria T. Arévalo
ISSN:2157-2526 JBTBD, an open access journal J Bioterror Biodef Bioterrorism: Infectious Diseases Vigilance is essential if we are to be prepared against biological threats stemming from bioterrorist attacks to emerging infectious diseases. Surveillance can provide valuable information for risk assessment and to make decisions on which pathogens to target with the most attention to development and stockpiling of preand postexposure prophylactic vaccines, drugs, or therapies. However, deciding on what probable bioweapon threats and emerging infectious diseases to invest time and resources on poses a challenge, especially during harsh economic times or in poorer regions. Should we thus, attempt to focus on the most plausible threat or the most devastating threat to occur even if it appears to be less probable?
如果我们要准备好应对新发传染病的生物恐怖袭击所带来的生物威胁,警惕是必不可少的。监测可以为风险评估提供有价值的信息,并决定针对哪些病原体,最重视开发和储存暴露前和暴露后预防性疫苗、药物或疗法。然而,决定在哪些可能的生物武器威胁和新出现的传染病上投入时间和资源是一项挑战,特别是在经济困难时期或在较贫穷地区。因此,我们是否应该试图把注意力集中在最可能发生的威胁或最具破坏性的威胁上,即使它看起来不太可能发生?
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引用次数: 1
Potential duel-use of bacteriophage related technologies in bioterrorism and biodefense. 噬菌体相关技术在生物恐怖主义和生物防御中的潜在双重用途。
Pub Date : 2012-11-23 DOI: 10.4172/2157-2526.1000121
Huahao Fan, Y. Tong
Bacteriophages are one of the simplest life forms on earth. They are natural killers of bacteria and exist in very large numbers in almost all natural habitats. With the ever-increasing threat posed by antibiotic-resistant bacteria, bacteriophages have regained the attention of biomedical scientists, but few reports have considered the use of bacteriophages in bioterrorism. This review describes the potential for bacteriophages to be used as bioterrorism agents or tools, as well as in counter-measures against bioterrorism.
噬菌体是地球上最简单的生命形式之一。它们是细菌的天然杀手,几乎在所有的自然栖息地都大量存在。随着耐药细菌的威胁日益增加,噬菌体重新受到生物医学科学家的关注,但很少有报道考虑将噬菌体用于生物恐怖主义。本文综述了噬菌体作为生物恐怖主义制剂或工具以及在生物恐怖主义对策方面的潜力。
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引用次数: 10
期刊
Journal of Bioterrorism and Biodefense
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