生物实验室安全

1、生物安全(nqun)防护FistXian Jiaotong University共四十一页实验室工作人员所处理的实验对象含有致病的微生物及其毒素时，通过在实验室设计建造、使用个体防护设置、严格遵从标准化的工作及操作程序和规程等方面采取综合措施，确保实验室工作人员不受实验对象侵染，确保周围环境不受其污染。 包括(boku)任何有潜在的感染可能的物质包括重组DNA研究什么是生物(shngw)防护共四十一页生物(shngw)安全事故举例1995年香港科技大学大四学生梁同学因吸入别的同学泼撒德酸酐而身亡1997年香港科技大学某访问学者因未按规定使用(shyng)通风橱而造成他人肺部伤害，该访问学者永不

2、被香港各大学欢迎2001年英国博布特莱尔实验室东北方向50公里的布伦特伍德地区发现口蹄疫疫情，后经调查发现该疫情暴发是口蹄疫病毒从实验室泄露导致的。2003年台北发现一位资深科学家感染SARS冠状病毒，该事故是由于处理包含SARS病毒标本时病毒泄露引起的，该类事件也在大陆和新加坡等地区发现。2007年，英国南部萨里郡发现口蹄疫事件，该事件也是实验室病毒泄露造成。DNA凝胶电泳EB染色过的琼脂糖凝胶不经处理，直接丢弃。工程菌培养液、实验室废液不经处理，直接倒入水池，重组的DNA进入土壤，可以存留40万年对工作人员的危害对环境的危害共四十一页理解危害物的生物学性质危害物在实验室之间的传播实验仪器以

3、及实验过程(guchng)中的危险性目标: 给实验者提供最有效的保护，减少实验过程中暴露的可能性 对所使用药品以及实验(shyn)环境风险的认定共四十一页实验室的分类(fn li)、分级分类：一般生物安全(nqun)防护实验室(不使用实验脊椎动物和昆虫)和实验脊椎动物生物安全防护实验室。分级：每类生物安全防护实验室根据所处理的微生物及其毒素的危害程度各分为四级。各级实验室的生物安全防护要求依次为：一级最低，四级最高。共四十一页一般生物安全(nqun)防护实验室-11.一级生物安全防护(fngh)实验室：实验室结构和设施、安全操作规程、安全设备适用于对健康成年人已知无致病作用的微生物，如用于教学

4、的普通微生物实验室等。 E. coli K-12 strains共四十一页2.二级生物安全(nqun)防护实验室 ：实验室结构和设施、安全操作规程、安全设备适用于对人或环境具有中等潜在危害的微生物。如：非呼吸道感染，非致病的物质共四十一页一般生物安全(nqun)防护实验室-33. 三级生物安全(nqun)防护实验室 实验室结构和设施、安全操作规程、安全设备适用于主要通过呼吸途径使人传染上严重的甚至是致死疾病的致病微生物及其毒素，通常已有预防传染的疫苗。 艾滋病病毒的研究(血清学实验除外)应在三级生物安全防护实验室中进行。 肺结核病毒，SARS等共四十一页一般(ybn)生物安全防护实验室-44.

5、 四级生物安全防护实验室 实验室结构和设施、安全操作规程、安全设备适用于对人体具有高度的危险性，通过气溶胶途径传播或传播途径不明，目前尚无有效的疫苗或治疗方法的致病微生物及其毒素。与上述情况类似的不明微生物，也必须在四级生物安全防护实验室中进行。待有充分数据(shj)后再决定此种微生物或毒素应在四级还是在较低级别的实验室中处理。FORGET ABOUT IT! 共四十一页一级生物(shngw)安全防护实验室的基本要求 1. 安全设备和个体防护 1.1 一般无须使用生物安全柜等专用安全设备。 1.2 工作人员在实验时应穿工作服，戴防护眼镜。 1.3 工作人员手上(shu shn)有皮肤破损或皮疹

6、时应戴手套。共四十一页一级生物安全防护(fngh)实验室的基本要求2 实验室设计和建造的特殊要求 2.1 每个实验室应设洗手池，宜设置在靠近出口处。 2.2 实验室围护结构内表面应易于清洁。地面应防滑、无缝隙，不得(bu de)铺设地毯。 2.3 实验台表面应不透水，耐腐蚀、耐热。 2.4 实验室中的家具应牢固。为易于清洁，各种家具和设备之间应保持生物废弃物容器的台(架)。 2.5 实验室如有可开启的窗户，应设置纱窗。 共四十一页生物(shngw)安全柜有效地减少身体和有毒物品的物理(wl)接触。高校过滤可以去除微粒 （最小 0.3 uM），效率达到99.97% 。 三种不同分级的安全柜，从而

7、提供不同的保护。共四十一页使用(shyng)生物安全柜使用之前：确保BSC被认证(rnzhng)过关闭 UV ；打开荧光灯工作区表面消毒工作之前5-10分钟开启鼓风机共四十一页WORKING SAFELY IN A BSC共四十一页After using the cabinet:Leave blower on at least 5 minutes to purge cabinetRemove and decontaminate equipment and materialsDisinfect cabinet surfacesTurn off blower and fluorescent lam

8、p, turn on UV lampWORKING SAFELY IN A BSC共四十一页Maintenance:Before and after each use - Work surfaces wiped downWeekly- UV lamp should be wiped cleanMonthly - All vertical surfaces wiped downAnnually- UV lamp intensity verified - Decontamination with formaldehyde gas (ORM)- Certification (ORM)WORKING

9、SAFELY IN A BSC共四十一页Bacterial:76% from clinical labs8% from research labsExposure:60% acquired from inhalationOther exposures include: digestion, sharps, splashes, direct and indirect contact Laboratory Acquired Infections (LAI)共四十一页Viral16% from clinical labs 70% from research labs32% from animal r

10、elated activities Laboratory Acquired Infections (LAI)共四十一页Biohazardous Waste CategoriesCultures and stocks of infectious agents and associated biologicalslaboratory wastebiological production wastediscarded live and attenuated vaccinesculture dishes and related materials共四十一页Biohazardous Waste Cate

11、goriesLiquid human and animal wasteliquid or semi-liquid blood and blood products and body fluidscontaminated items that would release blood or items that are caked with blood or other potentially infectious materials; NOT including urine or materials stained with blood or body fluidsinfectious anim

12、al waste (research)共四十一页Biohazardous Waste CategoriesPathological病理性的 wastetissuesbody parts other than teethproducts of conceptionfluids removed by trauma or during surgery or autopsy/necropsy or other medical procedure and not chemically fixed.共四十一页And More Biohazardous Waste CategoriesAnimal and

13、plant pathogen wasteRecombinant DNA wasteSharps共四十一页Biowaste vs. Trash3 basic questions to differentiate:Is it contaminated with viable biological material?Can blood or other regulated body or biological fluids be released?Is it a sharps hazard? 共四十一页#1Is it contaminated with viable biological mater

14、ial?Examples:Contaminated lab wastePersonal protective equipment used for handling potentially infectious materials (including handling infected animals or their products) Wastes from infectious disease research (carcasses, body fluids)共四十一页#2Can blood or other (regulated) body fluids or viable biol

15、ogical materials be released?Some ExamplesTubes of bloodVacuum flasks containing body fluids or cell line waste共四十一页Managing Liquid Biohazardous WasteStorage:Label and secure bulk vessels if not disposed of immediatelyTreatment:Chemical disinfection ORAutoclaveDisposal: THENFlush to sewer 共四十一页10% b

16、leach solution good for general disinfection High organics use 20% Needs to be made weeklyTest contact timeEthanolUse 70% solution (most effective)Longer contact time and flammable*Should research and know effectiveness and contact time for the best disinfectant against your agent! Disinfection 共四十一

17、页Managing Non-Sharp Biohazardous Wastelabeled container lined with a biohazardous waste bagequipped with a lid.共四十一页Managing Non-Sharp Biohazardous WasteSecurely tie bags for transport to treatment/collection site.When moving wastes, use secondary containment; avoid using public halls and elevators.

18、 共四十一页“Breakable” Non-sharps BiowasteStore in labeled containersthat are puncture-resistant, closable and will capture leakage, BUT.Do NOT use SHARPS containers!共四十一页Effective Waste AutoclavingLeave bag open during autoclaving or loosely closedAdd water to bag prior to autoclaving if primarily dry m

19、aterialsSteam must contact materialsPlace bag in autoclavable tray with sides共四十一页Treated Waste Bag DisposalAllow waste bag to coolUse fume hood to reduce odorsSecurely tie bag shutPlace bag in a non-transparent black bag for regular disposalRemember: NO ORANGE BAGS IN DUMPSTER!共四十一页#3Is it a sharps

20、 hazard?Examples:needlessyringesscalpelsall biologically contaminated objects that can easily penetrate skin (Pasteur pipettes,razor blades, etc.)Place sharps in approved sharps container for disposal!共四十一页Syringes in research settings should be disposed of as a sharp to avoid public relations conce

21、rns!共四十一页Sharps ContainersContainers must be leak-proof, puncture-resistant, closable & labeled with the biohazard symbol.Proper sharps containersmust be used forboth clinic andfield work.共四十一页Proper Use of Sharps ContainersPlace tops on containers before use on lab benchDont forget to date the container when first put into useRemember: sharps containers are aone-way disposal system共四十一页Proper Use of Sharps ContainersUse sharps containers for sharps ONLY!No solid biohazardou