实验室生物安全

1、Laboratory Biosafety 实验室生物安全,2020/6/25,OHS 4,2,Objectives目的,By the end of this session you will have an understanding of须了解: Classifications of biohazardous material危险生物材料分类 Risk group pathogens病原体危险性分组 definitions定义 Routes of exposure接触途径 Barriers for containment隔离设施 Methods of decontamination排除污染的

2、方法,2020/6/25,OHS 4,3,Definition定义,Biohazardous Materials 危险生物材料: Biological agents or substances present in, or arising from, the work environment which presents, or may present, a hazard to the health or well being of the worker or community存在、或产生于工作环境，可能导致群体或个人的健康危害的生物介质或物质,2020/6/25,OHS 4,4,Bioha

3、zardous Agents 危险生物介质,Classified into eight groups分为8组: cultured animal cells and the potentially infectious agents these cells may contain培养的动物细胞，并可能发生感染 primate body fluids and other potentially infected clinical specimens灵长类动物体液和可能传染的医用样品 microorganisms including Bacteria, Viruses, Fungi, Rickett

4、siae and Chlamydiae微生物类包括细菌、病毒、真菌、立克次氏体和Chlamydiae等,2020/6/25,OHS 4,5,Biohazardous Agents 危险生物介质,Parasites寄生虫 Allergens过敏原、 tissues from experimental animals including animal dander实验动物组织包括动物头屑 plant viruses, bacteria and fungi植物病毒、细菌和真菌 toxins (bacterial, animal or plant)生物毒素（细菌、动物或植物）,2020/6/25,OH

5、S 4,6,Laboratory Biosafety 实验室生物安全,Two main sources of Biohazardous material in use in most Laboratories实验室常用危险生物材料的两个主要来源: Pathogens used and tested for in Micro Dept微生物实验室试验用病原体 Biological fluid samples (eg. Plasma, urine), submitted for chemical analysis用于化学分析的生物液体样品（如血浆、尿）,2020/6/25,OHS 4,7,Labo

6、ratory Biosafety 实验室生物安全,The risk group of pathogens as classified by WHO世界卫生组织对病原体危险性分组 Risk Group 1 Agents 第一组介质 Low individual and community risk 低个体和群落风险 Risk Group 2 Agents第二组介质 Moderate individual risk, limited community risk中度个体风险，有限群落风险 Risk Group 3 Agents第三组介质 High individual risk, low comm

7、unity risk高个体风险，低群落风险 Risk Group 4 Agents第四组介质 High individual risk, high community risk高个体风险，高群落风险,2020/6/25,OHS 4,8,Risk Group 1 Agents 第一组介质,Non-pathogenic bacteria, fungi非病原体细菌、真菌 No special facilities, equipment or procedures are required无需特殊设备或操作 Standard well-designed experimental animal and

8、laboratory facilities and basic safe laboratory practices suffice标准的设计合理的实验动物、实验设备及满足基本安全试验操作 Hand-washing facilities must be provided必须提供洗手池 Disinfectants must be properly used必须合理使用消毒剂,2020/6/25,OHS 4,9,Risk Group 2 Agents 第二组介质,E. Coli, Salmonella, ringworm fungi, influenza virus, herpes simplex

9、virus大肠杆菌、沙门氏菌、 ringworm真菌、流行性感冒病毒和单纯疱疹病毒 Laboratory separated from other activities, biohazard sign, room surfaces impervious and readily cleanable实验室与其它活动分开，贴有危险生物标志，室内台面防渗，随时可进行清洁 Equipment should include an autoclave, certified HEPA filtered class I or II biological safety cabinet 实验设施包括高压灭菌锅、 高

10、效空气过滤器、一级或二级生物安全柜 All contaminated material to be properly decontaminated所有污染材料须经净化,2020/6/25,OHS 4,10,Risk Group 3 Agents 第三组介质,Anthrax, tuberculosis, HIV, hanta virus炭疽、结核、爱滋和hanta病毒 Specialized design and construction of laboratories, with controlled access double door entry and body shower实验室须专门

11、设计，双道门和淋浴 All wall penetrations must be sealed所有墙体渗漏必须进行密封 Ventilation system design must ensure that air pressure is negative to surrounding areas at all times, with no recirculation of air通风系统必须保证与周围区域一直呈负压，不可使空气再流通,2020/6/25,OHS 4,11,Risk Group 3 Agents 第三组介质,Air exhausted through a dedicated exh

12、aust or HEPA filtration system空气排放须经过专用排气系统或高效空气过滤器系统 Laboratory windows sealed and unbreakable实验室窗户须密闭并不可击碎 Backup power available应有备用发电机 Certified HEPA filtered class II biological safety cabinet经鉴定的高效空气过滤器二级生物安全柜 Dedicated handwashing sink with foot, knee or automatic controls, located near the e

13、xit在出口处设有专门的可用脚、膝盖或自动控制的洗手池,2020/6/25,OHS 4,12,Risk Group 4 Agents 第四组介质,Infectious agents are all viruses具有传染性的介质全部为病毒 Ebola viruses埃博拉病毒 Herpes B virus (Monkey virus) B 疱疹病毒 Foot and Mouth Virus脚和口病毒,Foot and Mouth Virus,2020/6/25,OHS 4,13,Risk Group 4 Agents 第四组介质,Highest level of containment最高级别

14、屏蔽 Represents an isolated unit that is completely self-contained to function independently设立一个设备齐全的隔离单元，完全能够独立操作 Facilities are highly specialized, secure with an air lock for entry and exit设备专门设计，由空气锁控制出入 Class III biological safety cabinets or positive pressure ventilated suits三级生物安全柜，正压通风 Separat

15、e ventilation system with full controls to contain contamination为全面控制污染物，采用单独的通风系统,2020/6/25,OHS 4,14,Routes of Exposure 接触途径,The most common routes of exposure are主要为: Oral口 Respiratory呼吸 Skin puncture皮肤划伤 Penetration through unbroken skin皮肤渗透 Conjunctival结膜 Vectors (mosquitoes, fleas, ticks)带菌者（蚊子

16、、跳蚤、虱子）,2020/6/25,OHS 4,15,Routes of Exposure 接触途径,Respiratory exposures account for 65-75% of all infections for which a cause is known.传染途径中有65-75%是呼吸造成的 It is therefore critical for people working with biohazardous and potentially infectious materials to know the risks associated with them and th

17、e appropriate laboratory techniques to prevent unnecessary exposures.因此实验操作中接触危险生物材料或潜在传染性材料时，了解其危害，采用适当技术防止不必要的接触十分关键,2020/6/25,OHS 4,16,Primary Barriers 第一道隔离,primary containment is the first line of defense when working with infectious organisms当工作中与传染性生物体接触时，第一道隔离是第一道防卫 glove boxes手套箱 biological

18、 safety cabinets生物安全柜 animal caging equipment动物笼养,2020/6/25,OHS 4,17,Primary Barriers 第一道隔离,The general principles applicable to containment within a primary barrier system include the following:第一道隔离一般原则 Minimize the volume to be contained.隔离体积最小化 Provide safe (i.e. non-contaminating) transfer of m

19、aterial into and out of the container without destroying the barrier.采用安全的从容器中取放材料的方法，勿毁坏隔离 Provide means for decontamination of the enclosure and effluents.采用净化隔离栏和污物的方法,2020/6/25,OHS 4,18,Biological Safety Cabinets 生物安全柜,Three types分为3种 Class I 一级 protects worker and environment保护操作者和环境 Class II二级

20、 protects worker, environment and product保护操作者、环境和产品 Class III 三级 for Group 4 work, maximum protection of worker, environment and product用于第4类危险品的操作，最大限度保护操作者、环境和产品,2020/6/25,OHS 4,19,Class I BSC一级生物安全柜,Similar to chemical fumehood except exhaust passes through HEPA filter类似化学实验室的通风橱，排出的气体要通过高效空气过滤器

21、 Often used to enclose equipment such as centrifuges or balances常常用于放置离心机和天平,2020/6/25,OHS 4,20,Class I BSC Schematic 一级生物安全柜示意图,2020/6/25,OHS 4,21,Class II BSC 二级生物安全柜,Supplies a downward flow of HEPA filtered air to protect product提供气流向下的经高效空气过滤器净化的空气，保护产品 Further classified based on ratio of exha

22、usted vs. re-circulated air进一步基于排气/再循环空气比值进行细化为A、B1、B2和B3型 Type A 75 fpm face velocity, 30% internal exhaust Type B1 100 fpm, 50% external exhaust Type B2 100 fpm, 100% external exhaust Type B3 100 fpm, 30% external exhaust,2020/6/25,OHS 4,22,Class II BSC二级生物安全柜,Type A,Type B1,The cabinet exhaust ne

23、eds to be connected to the building exhaust.,2020/6/25,OHS 4,23,Class III BSC二级生物安全柜,For Risk Group 4 materials第四组介质 Sealed unit 密闭单元 “glove box” “手套箱” Exhaust is passed through double HEPA filters or otherwise treated (e.g. air incineration) to prevent escape of level 4 material to the environment排

24、气须经过双层的高效空气过滤器或其它处理方法（如空气焚烧），阻止第四组介质进入环境,2020/6/25,OHS 4,24,Class III BSC二级生物安全柜,glove ports with O-ring for attaching arm-length gloves to cabinet圆形手套固定端口， 可戴手臂长度的手套在安全柜中操作 B. sash窗框 C. exhaust HEPA filter 排气滤膜 D. supply HEPA filter 供气滤膜 E. double-ended autoclave or pass-through box双头高压灭菌锅或贯通箱,2020

25、/6/25,OHS 4,25,Secondary Barriers第二道隔离,If primary barriers fail第一道隔离失败 clothing and other items of personal protective equipment become an important line of defense 工作服及其它个人防护用品成为重要的防线,2020/6/25,OHS 4,26,Decontamination净化,Sterilization杀菌 Complete inactivation of the infective potential of a biologic

26、al agent完全阻止生物介质的可能传染 Usually by autoclave常用高压灭菌锅 Disinfection消毒 Reduction in number or viability of pathogenic microorganisms below the point of being able to transmit disease减少微生物病原体的数量和繁殖，使其无法传播疾病,2020/6/25,OHS 4,27,Sterilization杀菌,By steam at a temp. of 121C. for 15 minutes 121C蒸15分钟 By dry heat

27、 at a temperature of 160-170 C. for 2-4 hours干燥加热需160-170 2-4 小时,2020/6/25,OHS 4,28,Disinfection消毒,Mode of action Cell lysis 细胞溶解 Protein coagulation or denaturation 蛋白凝固或变性 Enzyme denaturation or inactivation酶变性或失活 Destruction of enzyme substrate酶作用物破坏,2020/6/25,OHS 4,29,Disinfection消毒,Many chemica

28、l disinfectants available主要化学消毒剂 Alcohols醇类 Formaldehydes甲醛 Phenolic Compounds酚化合物 Quaternary Ammonium Compounds or Quats季铵盐 Chlorine Compounds氯化物 Iodine Compounds碘化物 Glutaraldehydes戊二醛 Hydrogen peroxides过氧化氢 Chlohexidine compounds六氯化合物,2020/6/25,OHS 4,30,Alcohols醇类,Ethanol or iso-propanol乙醇或丙醇 Dena

29、ture proteins蛋白质变性 Effective against lipid containing viruses, enveloped viruses and vegetative bacteria对含油脂病毒、膜包被病毒和植物细菌有效 Less effective against mycobacterium and fungi对分支杆菌和真菌作用小 Flammable, inactivated by organic matter, volatile可燃、遇有机物活性差、挥发,2020/6/25,OHS 4,31,Aldehydes醛类,Formaldehyde and glutar

30、aldehyde蚁醛和戊二醛 Effective against a broad spectrum of microorganisms对广谱微生物有效 Requires long (20-30 min) contact time作用时间长（20-30分钟） Less susceptible to inactivation by organic material对有机体的去活作用小 Short use-life使用有效期短 Human health concerns - respiratory irritant, contact dermatitis对人体健康有害-呼吸刺激、接触性皮炎,2020

31、/6/25,OHS 4,32,Phenolics酚类,Usually phenolic compound with detergent常为分化合物清洁剂 Effective against vegetative bacteria, fungi and lipid-containing viruses对植物细菌、真菌和含脂病毒有效 Effective over a wide pH range pH 范围宽 Skin and eye irritation皮肤和眼的刺激 Not effective against spores对孢子无效,2020/6/25,OHS 4,33,Quaternary A

32、mmonia Componds 季铵盐化合物,Cationic detergents阳离子清洁剂 / strong surfactants强表面活性剂 Effective against vegetative bacteria (some) and non-lipid-containing viruses对植物细菌（部分）和不含脂病毒有效 Stable, Non-irritating稳定、无刺激 Ineffective against mycobacteria, lipid-containing viruses, spores对分支杆菌、含脂病毒和孢子无效 Neutralized by org

33、anic material (esp. proteins)与有机物（特别是蛋白）中和,2020/6/25,OHS 4,34,Chlorine Componds氯化物,Strong oxidant强氧化剂 “Universal” decontaminant普遍采用的纯化剂 May be prepared as 1/100 or 1/50 (for spores and mycobacteria) dilution from household bleach and tap water用家用漂白剂或自来水稀释制备成1/100 或 1/50 （对孢子和分枝杆菌）溶液 Dilute solutions

34、 are unstable, prepare frequently稀释溶液不稳定，常配 Neutralized by organic material用有机物质中和 Corrosive具腐蚀性,2020/6/25,OHS 4,35,Iodine Componds碘化物,Similar in action to chlorine与氯相似 Effective against vegetative bacteria and viruses对植物细菌和病毒有效 Less corrosive and toxic than chlorine比氯的腐蚀性和毒性低 Self-indicating自显示: brown棕-yellow黄-clear无色 May stain surfaces使表面染色 Neutr