狂犬病毒的实验在多数情况下可在 生物安全2级实验室 进行,仅在少数特殊情况下才需要在 生物安全3级实验室 进行。 对狂犬病的实验研究,一般情况下可在 生物安全 2 级实验室 进行,在某些情况下,可考虑在采用更高标准的 生物安全 3 级实验室 进行。 世界卫生组织( WHO ) 狂犬病专家磋商会第二次报告( 2013 年版)中关于狂犬病实验的生物安全问题是这样写的: “狂犬病是目前已经认识到的所有传染病中病死率最高的。因此,安全性对于从事狂犬病病毒的工作至关重要。一般来说, 生物安全2级实验室 的安全操作适用于动物处理、尸体解剖、样本采集、准备、处理等常规的实验室活动。除了符合基本设施要求之外,预防措施还包括个人防护设备(如服装、手套、护目镜) 和疫苗接种。 在某些情况下,如生产大量浓缩病毒、进行可能产生气溶胶的操作(例如组织悬液匀化)、操作尚不清楚当前的预防措施是否具有保护效果的新分离狂犬病病毒时,可考虑采用 生物安全3级 。所有用于操作传染性病原的国家安全指南均应严格遵守。” 既然狂犬病的病死率是所有传染病中最高的(发病后死亡率 10 0%),为什么 狂犬病毒的实验在多数情况下可在生物安全 2级实验室进行,仅在少数特殊情况下才需要在 生物安全 3 级实验室 进行 呢? 这是由于狂犬病毒在环境中容易失活(死亡),不易传播(传播方式很特殊,通常只能通过疯狗咬伤才能传播,原则上不能通过空气和接触传播),而且早在 100 多年前,就已有 100% 有效的疫苗可进行针对性的预防。 狂犬病毒是一种已经得到充分研究的病毒,人类对它已有充分的认识,已有控制甚至彻底消除狂犬病的成熟的技术手段。事实上,目前在全球所有发达国家和部分发展中国家,都已经基本消灭了狂犬病(人被狗咬了也不用打疫苗,到一定时候甚至连狗也不用打疫苗)。在中国,只要能正确运用现有的有关狂犬病的知识,其实狂犬病并不那么可怕,狂犬病相对而言是比较容易预防和控制的。在中国,狂犬病也有希望在不久的将来被彻底消除。 为了能 更全面地理解 狂犬病毒的相对危险性 ,有必要对 实验室生物安全防护水平分级 的基本知识进行介绍。 世界卫生组织( WHO )一直非常重视生物实验室安全问题,新修订的 《实验室生物安全手册》于 1983 年出 版 。 中国也于2008年颁布了新修订的国家标准《实验室生物安全通用要求》(GB 19489)。 传染性微生物根据其致病能力和传染的危险程度等可划分为四类(四级); 第一级的微生物与人类成人的健康和疾病无关; 第二级在人类所引起的疾病很少是严重的,而且通常有预防及治疗的方法; 第三级在人类可以引起严重或致死的疾病,可能有预防和治疗的方法; 第四级在人类可以引起严重或致死的疾病,而且通常无预防和治疗的方法(如炭疽杆菌、霍乱弧菌、埃博拉病毒、 马尔堡病毒、拉萨( Lassa )病毒、克里米亚 - 刚果出血热病毒、 天花病毒等)。 根据对所操作生物因子采取的防护措施,将实验室 生物安全防护水平 (Biosafety Level,BSL) 分为 P1 、 P2 、 P3 和 P4 共四个生物安全等级。 “P” 是英文 Protection (保护)的缩写。生物安全 4 级( BSL 4 )实验室,又称 P4 实验室,是生物安全最高等级的实验室,可有效阻止最危险的传染性病原体释放到环境中,同时为研究人员提供安全保障。 生物安全一级(P1)实验室 因主要从事通常不会引起人类或者动物致病的微生物的操作,因而,生物风险十分有限。但是,对一些特殊人群,如孕妇、婴幼儿、过敏体质或有特定疾病的人员,仍可能存在较大的风险。此外,还应从环境安全、实验结果的质量等角度考虑对微生物污染进行控制。P1实验室的很多活动是培训和教学,当涉及学生教学时,由于学生安全意识差、人员多,可能会有一些意想不到的状况发生。 生物安全二级(P2)实验室 操作对人类或者动物致病的微生物,有生物风险。而且,这类实验室众多、工作量大、工作种类多、样本复杂、从业人员多、未知因素多、安全意识相对弱,是生物安全事故发生概率最高的一类实验室。风险等级为2级的病原,可对个体产生不同形式和不同程度的损害,甚至导致死亡。由于样本复杂,可能包含未知病原或高风险等级的病原,有导致严重后果的潜在危险,包括在个体之间的传播。 风险等级为3级之微生物的重要特征是传播性强、危害大,是高致病性微生物。一旦发生感染事件,可能严重危害人员的健康和生命,引起社会恐慌,造成巨大损失,2004年发生在北京的实验室SARS病毒感染事件就是典型的案例。 从实验室活动看, P3实验室 和P2实验室并无本质的区别,但人员的心理压力会较高、操作灵活性降低(安全防护要求和心理压力均是影响因素)。从操作对象看,P3实验室风险级别更高,但样品来源和目标微生物通常比较确切,从设施设备、个体防护、人员、管理体系等方面的要求看,都显著高于P2实验室。风险等级为3级的病原微生物,绝大多数是通过气溶胶经呼吸道造成人员感染,因而,防护难度相对较大。 风险等级为4级之微生物是目前人类已认识或尚未认识的最危险的病原微生物,传播性强、感染后死亡率高。比如天花病毒,在自然界中存活力强、易于通过气溶胶传播、毒力高,曾给人类带来极大的灾难。由于生物风险极高,因而要求必须保证人员与操作对象在完全隔离的状态下从事相关工作。 进入 P4 实验室 的研究人员都必须换上隔离正压防护服, 即在人与微生物之间设置可靠的隔离操作系统。为保证环境安全,须采用两层HEPA过滤器处理排出的气体,所有废弃物须经可靠消毒后才能移出实验室。 P4 实验室有严格、复杂的管理程序,为了保证 P4 实验室的绝对安全,只有得到批准和持有磁卡通行证的人才能进入,有的通过指纹门禁系统进入,而且所有出入的人员都由电脑记录在案。 目前 P4 实验室 在全球的数量和分布 P4 实验室 是生物安全顶级实验室,这不仅仅是指它在生物安全方面是顶级的,其造价、运营和维护也是最贵的。因此,P4实验室目前在全球范围内数量很少,总共只有约50个,这其中还包括一些正在建设中的或仅断续运转的(相关详情请参看本文所附表格)。 2015 年 01 月 31 日,经过 中国科学院武汉病毒研究所 和 法国 的相关科研人员十多年的努力, 中国科学院 武汉国家生物安全实验室 ( 武汉 P4 实验室 ) 在武汉建成,标志着中国正式拥有了研究和利用烈性病原体的硬件条件。 当前,除中国外,全球有约 20 个国家或地区拥有 P4 实验室(包括法国、加拿大、德国、澳大利亚、美国、英国、加蓬(法国巴斯德所)、瑞典、南非、印度、 阿根廷、捷克、匈牙利、意大利、日本、俄罗斯、瑞士、中国台湾等)。 这总共约 50 个 P4 实验室遍布世界五大洲,其中非洲大陆只有两个(分别位于南非和加蓬)。单就国家而论,美国的 P4 实验室最多,共有 15 个(包括正在建设中的)。相关 详情可参考下表(资料来源: https://en.wikipedia.org/wiki/Biosafety_level#Biosafety_level_4 ) 。 The following is a list of exis ting BSL-4 facilities worldwide(目前全球P4实验室列表): 序号 名称 所在地(国家或地区) 建成时间 说明 1 National Service of Healthcare and Agriculture Quality (SENASA) Buenos Aires , Argentina Diagnostic laboratory for Foot-and-mouth disease . 2 University of Melbourne – Doherty Institute for Infection and Immunity Melbourne , Victoria , Australia 2014 Diagnostic reference lab. 3 Australian Animal Health Laboratory Geelong , Victoria , Australia 1985 Large diagnostic and research lab equipped with two BSL-4 animal rooms. 4 National High Security Laboratory Victoria , North Melbourne , Australia Operates under the auspice of the Victoria Infectious Diseases Reference Laboratory. 5 National Microbiology Laboratory Winnipeg , Manitoba , Canada Located at the Canadian Science Centre for Human and Animal Health , it is jointly operated by the Public Health Agency of Canada and the Canadian Food Inspection Agency . 6 Wuhan Institute of Virology of the Chinese Academy of Sciences Wuhan , Hubei , China 2015 Wuhan Institute of Virology has existed since 1956 and already hosted BSL3 laboratories. A BSL4 facility was completed in 2015, and became the first BSL-4 laboratory in China. 7 Biological Defense Center Těchonín , Pardubice , Czech Republic 1971, rebuilt 2003-2007 Located at the Centrum biologické ochrany (Biological Defense Center) 8 Jean Mérieux BSL-4 Laboratory Lyon , Rhône-Alpes , France 1999 Built and owned by the Fondation Mérieux . Since 2004, operated by INSERM . 9 Laboratoire de la DGA Essonne , Vert-le-Petit , France 2013 Operated by the Ministry of Defense. 10 Centre International de Recherches Médicales de Franceville Gabon This facility is operated by a research organization supported by both Gabonese (mainly) and French governments, and is West Africa's only P4 lab (BSL-4). 11 Robert Koch Institute Berlin , Germany 2015 Diagnostic and experimental lab facility. 12 Bernhard Nocht Institute for Tropical Medicine Hamburg , Germany Part of the Leibniz Center Infection . National reference lab for tropical viruses. 13 Friedrich Loeffler Institute Germany , Isle of Riems ( Greifswald ) 2010 Focus on animal viral diseases and diagnostics. 14 Philipps University of Marburg Marburg , Germany 2008 Focuses on hemorrhagic fever viruses. 15 Országos Epidemiológiai Központ – National Center for Epidemiology Budapest , Hungary 1998 Division of Virology operates three WHO National Reference Laboratories. The BSL-4 biosafety laboratory provides a modern means to process dangerous imported zoonotic viral pathogens. 16 High Security Animal Disease Laboratory (HSADL) Bhopal , India 1998 This facility deals especially to zoonotic organisms and emerging infectious disease threats. 17 Centre for Cellular and Molecular Biology Hyderabad , India 2009 National BSL-4 Containment Facility for Human Infectious Diseases. 18 Microbial Containment Complex Pune , India 2012 Run by the National Institute of Virology . 19 Istituto Nazionale per le Malattie Infettive Rome , Lazio , Italy 1997 The National Institute of Infectious Diseases used to operate within the Lazzaro Spallanzani hospital; the facility is now independent and is home to five BSL-3 labs as well as a single BSL-4 laboratory, which was completed in 1997. 20 National Institute for Infectious Diseases Musashimurayama , Tokyo , Japan 2015 Located at National Institute for Infectious Diseases, Department of Virology I. Built in 1981; operated at BSL-3 until 2015 due to opposition from nearby residents. 21 Institute of Physical and Chemical Research (RIKEN) Tsukuba , Ibaraki , Japan 1984 Facility completed in 1984 but not operated as BSL-4 due to local opposition. State Research Center of Virology and Biotechnology (VECTOR) Koltsovo , Novosibirsk Oblast , Russia One of two WHO -approved facilities for work on variola virus . 22 National Institute for Communicable Diseases Johannesburg , South Africa 2002 23 Public Health Agency of Sweden Solna , Sweden 2001 The only BSL-4 facility in the Nordic region . Constructed for research and diagnostics of hemorrhagic fever viruses . 24 University Hospital of Geneva Switzerland Glove box type laboratory; primarily for handling clinical samples. 25 Spiez Laboratory Spiez , Switzerland 2013 Run by Federal Office for Civil Protection and the Federal Department of Defence, Civil Protection and Sports . 26 Institue of Medical Virology Zurich , Switzerland Part of the University of Zurich . Primary purpose is diagnostics of highly pathogenic viruses. 27 Institute of Preventive Medicine Taipei National Defense University , Taipei , Taiwan 28 Health Protection Agency 's Centre for Infections Colindale , United Kingdom Department of Health laboratory. Diagnostics for various viral diseases. Part of the European Network of Biosafety-Level-4 Laboratories. 29 National Institute for Medical Research London , United Kingdom Medical Research Council laboratory. Research and diagnostics for highly pathogenic viruses. 30 Institute for Animal Health Pirbright , United Kingdom Biotechnology and Biological Sciences Research Council laboratory. Research on highly pathogenic animal diseases. 31 Defence Science and Technology Laboratory Porton Down , United Kingdom Ministry of Defense laboratory. Focuses on protection from biological weapons. 32 Centre for Emergency Preparedness and Response Porton Down , United Kingdom Department of Health laboratory. Diagnostics and research for haemorrhagic fever viruses. Part of the European Network of Biosafety-Level-4 Laboratories. 33 Francis Crick Institute London , United Kingdom 2015 Has BSL-4 space but does not work on human pathogens. 34 National Institute for Biological Standards and Control Potters Bar , Hertfordshire , United Kingdom Department of Health and Home Office laboratory. Develop assays and reagents for research on virulent pathogens. 35 Veterinary Laboratories Agency Addlestone , Surrey , United Kingdom Department for Environment, Food and Rural Affairs laboratory. Diagnostics and research for animal diseases. 36 Merial Animal Health Pirbright , Surrey , United Kingdom Private lab. Produces vaccines against foot and mouth disease and bluetongue disease . 37 Centers for Disease Control and Prevention Atlanta , Georgia , United States Currently operates in two buildings. One of two facilities in the world that officially hold smallpox . 38 Georgia State University Atlanta , Georgia , United States 1997 Research focus on B virus . 39 National Bio and Agro-Defense Facility (NBAF), Kansas State University Manhattan , Kansas , United States 2022 (expected) Under construction. Facility to be operated by the Department of Homeland Security , and replace the Plum Island Animal Disease Center . Expected to be operational by 2022-2023. 40 National Institutes of Health (NIH) Bethesda , Maryland , United States Located on the NIH Campus, it currently only operates with BSL-3 agents. 41 Integrated Research Facility Fort Detrick , Maryland , United States Operated by NIAID . Focuses on animal models of human diseases. 42 National Biodefense Analysis and Countermeasures Center Fort Detrick , Maryland , United States Operated by the Department of Homeland Security . Focus on potential bioterrorism threats. 43 US Army Medical Research Institute of Infectious Diseases (USAMRIID) Fort Detrick , Maryland , United States 1969 Run by the U.S. Army . Research focuses on biological threats to the U.S. military. 44 National Emerging Infectious Diseases Laboratory (NEIDL), Boston University Boston , Massachusetts , United States Focus on potential threats to public health. 45 Rocky Mountain Laboratories Hamilton , Montana , United States 1928 NIAID laboratory. Focus on vector-borne diseases. 46 Galveston National Laboratory , National Biocontainment Facility Galveston , Texas , United States Opened in 2008, facility is operated by the University of Texas Medical Branch . 47 Shope Laboratory Galveston , Texas , United States 2004 Operated by the University of Texas Medical Branch . 48 Texas Biomedical Research Institute San Antonio , Texas , United States 1999 The only privately owned BSL-4 lab in the US.