appendix a - biosafety in the virology laboratory

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Appendix A: Biosafety in thevirology laboratoryWorking with viruses which are infectious tohumans poses a considerable r isk of labora-tory-acquired infect ion. This was demon-strated on numerous occasions in the past,and led to the establ ishment of a classif ica-t ion of viruses and other infectious agentsinto four groups depending upon the hazardsto the laboratory worker. A ful l descript ion ofthe histor ical development and current guide-l ines for biosafety can be found in Biosafety inMicrobiological and Biomedical Laboratoriesedi ted by JW Richmond and RW McKinney,third edit ion, May 1993. Published by the USDepartment of Health and Human Services,Publication number (CDC) 93-8395.

Here we reprint from that publication thesummary descr ipt ion of recommended safetypractices at each of the four levels (Table 1).

Biosafety level 4 (maximum containment)laboratories are expensive to maintain andexist only in Australia, Canada, France, Ger-many, Russia, South Afr ica, the UK and theUSA.

For human blood-borne pathogens, bio-safety precautions have been issued to healthcare and cl inical laboratory workers under theterm 'Universal Precautions' and these need tobe observed for al l work involving contact withpotential ly infectious blood and body f luids inaddit ion to ut i l izat ion of the appropriate biosaf-ety level laboratory. A full description of uni-versal precautions has been published.(Occupational Exposure to Bloodborne Patho-gens, Final Rule, US Federal Register 56,64175-64182, 1991).

Since the eradication of smallpox, routinevaccination with vaccinia virus has ceasedthroughout the wor ld. However work wi th vac-

cinia virus continues in many virology labora-tories because of i ts value as a gene vector,e.g. in various vaccinia-based expression sys-tems. In persons vaccinated within the pre-vious ten years, work with vaccinia virusposes no special r isk, and the US policy isthat such work can be conducted at BiosafetyLevel 2 by appropriately vaccinated persons ina laboratory area with appropriate warningsigns to prevent entry of unvaccinated per-sons. However, vaccination i tself can be asso-ciated with signif icant adverse reactions, andin the UK and other European countr ies vacci-nation of laboratory workers is not advised(and is il legal in some countries, e.g. Ger-many). The danger to unvaccinated laboratoryworkers who use vaccinia virus is that theycould become infected at an inappropr iatesite (e.g. the eye), with the possibility of aserious adverse reaction. The laboratory direc-tor may wish, therefore, to raise work withvaccinia virus to Biosafety Level 3, wheresuch unvaccinated persons are involved.

The fol lowing classif icat ion of vertebrateviruses potential ly infectious for humans indi-cates the appropriate biosafety level (2, 3 or 4)at which work should be per formed. Note thatsome vi ruses appear twice, depending uponthe type of work being undertaken. In gen-eral, work involving virus concentrat ion tohigh t i tre, production of aerosols, or virusinfection of an animal host may require ahigher biosafety level than that needed forother types of work. The laboratory director isresponsible for appropriate r isk assessment,and on occasion may decide to select a bio-safety level higher than that recommended inthe fol lowing classificat ion.

Virology Methods ManualISBN 0-12-46533 0-8 Copyright 9 1996 Academic P ress LtdAll rights of reproduction in any form reserved


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Virology methods manualLevel TwoAabahoyoAbrasAbu HammadAcadoAcaraAdeno-associated virus

BotambiBotekeBoubouiBuffalopoxBujaruBunyamweraBunyip

Eastern equ ine encephalitis tEdge HillEntebbe batEnterovirusesEpizootic haemorrhagicdisease (of deer)Epstein-BarrAdenovirusesAguacateAlfuyAlmpiwarAmapariAnanindeuaAnhangaAnhembiAnopheles A

Anopheles BApeuApoiArideArkonamAroaAruacArumowotAstrovirusAuraAvalonB19BagazaBahigBakauBakuBandiaBangoranBanguiBanziBarmah ForestBarurBataiBatamaBaulineBebaruBelmontBenevidesBenficaBetiogaBimitiBiraoBluetongueBoraceia358

Burg E ArabBushbushBussuquaraButtonwillowBwambaCacaoCache valleyCalifornia encephalitisCalmitoCalovoCandiruCape WrathCapimCaraparuCarey IslandCatuChacoChagresChandipuraChanguinolaCharlevilleChenudaChikungunya (vs 131/25)*ChilibreChobar GorgeCIo MorColorado tick feverCoronavirusCorripartaCotiaCowbone RidgeCowpoxCoxsackieCsiro VillageCuiabaCytomegalovirusD'AguilarDakar BatDengue ItDengue 2Dengue 3Dengue 4tDera Ghazi Khan

ErveEubenangeeEyachFlandersFort MorganFrijolesGamboaGan GanGomokaGossasGrand ArbaudGreat IslandGuajaraGuamaGumbo LimboHart ParkHazaraHepatitis AHepatitis BHepatitis CHepatitis DHepatitis EHepatitis FHepatitis GHighlands JHuachoHughesHuman herpesvirusHuman papillomavirusesIcoaracileriIleshaIlheusInfluenza AInfluenza BInfluenza CIngwavumaInkooIppyIrituiaIsfahanItaporangaItaqui

A p p e n d i x A


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A p p e n d i x AJamestow n Canyon Lone Star NyandoJapanaut Lukuni O'nyon g nyongJerry Slough Lymph ocytic choriomeningitis OkhotskiyJohnston e Atoll M'po ko OkolaJoinjakaka Madrid OlifantsvleiJuan Diaz Maguari OliverosJugra Mahogany Hammock OrfJunin (vs Cand id #1) Main Drain OribocaJurona Malakal OssaJutiapa Manawa PacoraKadam Manzanilla PacuiKaeng Khoi Mapp utta PahayokeeKaikalur Maprik PalyamKaisodi Marco ParainfluenzaKamese Marituba ParanaKammavan pettai Marrakai PataKannaman galam Matariya Pathum ThaniKao Shuan Matruh PatoisKarimabad Matucare Phnom-P enh batKarshi Measles P chi ndeKasba Melao PixunaKemerovo Mermet PointKern Canyon Milkers nodule PoliovirusKetapa ng Mi natitlan Pon golaKeterah Minnal PontevesKeuraliba Mirim PrecariousKeys tone Mitchell River PretoriaKismayo Modo c Prospect HillKlamath Moju PuchongKokobera Molluscum contagiosum Punta SalinasKolongo Mono Lake Punta ToroKoongol Montana myotis QalyubKotonkan leukoencepha litis QuaranfilKowanya ma Moriche Rabies virusK u n i n M osq ueiro ReovirusesKununurra Mossuril Respiratory syncytialKwatta Mount Elgon bat RestanLa Crosse Mum ps Retroviruses, including humanLa Joya Murutucu and simian immun odeficiencyLagos bat Mykines viruses (HIV-1, HIV-2, and SIV)Landjia Navarro RhinovirusesLangat Nepuyo Rift Valley fever (vs MP-12)Lanjan Ngaingan Rio BravoLas Maloyas Nique Rio GrandeLatino Nkolbisson Ross RiverLe Dantec Nola RotavirusesLebombo Norwalk Royal farmLednice Ntaya RubellaLipovnik Nugget SaboLokern Nyamanini SaboyaA p p e n d i x A : B i o s a f e t y in t h e v i r o lo g y l a b o r a t o r y 359


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V i r o l ogy m e t hods m anua lSaint FlorisSakhalinSalehabadSan AngeloSandfly fever (Sicilian)Sandfly fever (Naples)SandjimbaSangoSathuperiSawgrassSebokeleSeletarSembalamSerra do NavioShamondaShark RiverShuniSilverwaterSimbuSimian hemorrhagic feverSindbisSixgun CitySnow MountainSnowshoe HareSokulukSoldadoSororocaSpumavirusStratfordSunday CanyonTacaiumaTacaribeTaggertTahynaTamiamiTanapoxTangaTanjong RabokTataguineTehranTembeTembusuTensawTeteTettnangThimiriThottapalayamTibrogarganTimboTimboteua

TindholmurTorovirusToscanaToureTransmissible spongiformencephalopathiest(Creutzfeldt-Jakob, kuru,scrapie and re lated agents)TribecTrinitiTrivittatusTrubanamanTsuruseTurlockTyuleniyUganda S

UmatillaUmbreUnaUpoluUrucuriUsutuUukuniemiVaccinia (vaccinated persons)Varicella-zosterVelloreVenezuelan equine encephalo-myelitis (vs TC-83) * tVenkatapuramVesicular sto ma titis - IndianaVesicular stomatitis - NewJerseyVincesVirgin RiverWad MedaniWallaWanowrieWarrego

Western equine encephalitisWhataroaWitwatersrandWongaWongorrWyeomyiaYabapoxYaquinea HeadYatapox*Yellow fever (vs 17-D)YogueZaliv Terpeniya

ZeglaZikaZingilamoZirqaLevel ThreeAdelaide RiverAgua PretaAinoAkabaneAlenquerAlmeirimAltamiraAndasibeAntequeraAraguariAransas BayArbiaArboledasBabankiBatkenBelemBerrimahBhanjaBimboBlack Cr eek CanalBobayaBobiaBozoBuenaventuraCabassueCacipacoreCalchaquiCananeiaCanindeChikungunyaChimCoastal PlainsCocalConnecticutCorfuDabakalaDhoriDobrava-BelgradeDouglasDugbeEnseadaEstero RealEvergladesFlexal

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A p p e n d i x AFomede Mucam bo Santa RosaForecariah Munguba SantaremFort Sherman Murray Valley encephalitis SaracaGabek Forest Nairobi sheep disease Saumarez ReefGadgets Gully Naranjal SedlecGarba Nariva Sem liki ForestGermiston Nasoule Sena MadureiraGetah Ndelle SeoulGordil Ndumu Sep ikHantaan Negis hi Sho kweHerpesvirus simiae (B virus) New Minto Sin NombreHum an polyomaviruses (JC, Ngari SlovakiaBK) Ngoupe Somonelaco Nodamura SpipurIbaraki North way S po ndwen iIfe Odrenisrou St Louis encephalitisIngangapi Omo TaiInini Oriximina TamdyIsrael Turkey meningitis Oropouche Telok Forestissyk- kul Oru ngo Termei IItaituba Ouango ThiaforaItimirim Oubangui ThogotoItupiranga Oubi TilligerryJacareacanga Ourem TinarooJamanxi Palestina Tlacota lpanJapanese encephalitis Para TocioJari Paramushir TonateJunin Paroo River TtingaKairi Peaton TurunaKedougou Perinet Venezuelan equineKhasan Pery encephalitisKimberley Petevo Vesicular stom atitis (infectedKindia Picola animals)Koutango Piry WesselbronKyzylagach Playas West NileLake Clarendon Powassan XiburemaLlano Seco Pueblo Viejo YacaabaLoup ing III Purus YaoundeLymphocytic choriomeningitis Puu ma la Yellow fever(infected animals) Radi YokaMacaua Razdan Yug BogkanovacMapuera ResistenciaMayaro Rift Valley fever Level FourM boke Rochambeau AbsettarovMeaban Rocio Black Cree k Can al (infectedM ddleburg Sagiyama animals)Mobala Sal Vieja Congo-Crimean hemorrhagicMojui Dos Com pos Salanga feverMonte Dourado San Perlita EbolaMopeia San Juan GuanaritoAp pen d ix A : B iosa fe ty in the v i ro logy l abora to ry 361


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V i r o lo g y m e t h o d s m a n u a lHanzalova Machupo Russian Spring-SummerHypr Marburg encep halitisJunin Omsk hemorrhagic fever SabiaKum linge Rift Valley Fever (infected Sin Nom bre (infected animals)Kyasanur Forest disease animals) Tick-b orne enceph alitisLassa* Vaccine Straint Viruses marked with this dagg er are classified in Europe either one level above (for Level Two) orbelow (for L evel Three) the US levels.

BiosafetyLevel

T a b l e 1 S u m m a r y o f r e c o m m e n d e d b i o s a f e t y l e ve l s f o r i n f e c t io u s a g e n t sAgents Practices Safety Equipment Facilities (Secondary(Primary barriers) Barriers)Not known to cause Standard None required BSL-1 plus:disease in healthy Microbiological autoclave availableadults Pract icesAssociated with humandisease, hazard=auto-inoculation, ingestion,mucous membrane

Indigenous or exot icagents with potent ial foraerosol t ransmission;disease may haveserious or lethalconsequences

Dangerous/exot icagents which pose highrisk of life-threateningdisease, aerosol-t ransmit ted labinfections; or relatedagents with unknownrisk of t ransmission

BCL-1 practice plus:9 Limited access9 Biohazard warningsigns9 Biosafety manualdef ining any neededwaste decontaminat ionor medical surveil lancepolicies

BSL-2 pract ice plus:9 Controlled access9 Decontaminat ion ofall waste9 Decontaminat ion oflab clothing beforelaundering9 Baseline serum

BSL-3 pract ices plus:9 Clothing changebefore entering9 Shower on exit9 All materialdecontaminated on exitfrom facility

Primary barriers= ClassI or ii BSCs or otherphysical containmentdevices used for allmanipulat ions of agentsthat cause splashes oraerosols of infect iousmaterials; PPEs;laboratory coats;gloves; face protect ionas neededPrimary barriers= Classi or II BSCs or otherphysical containmentdevices used for allmanipulat ions ofagents; PPEs:protect ive lab clothing;gloves; respiratoryprotect ion as neededPrimary barriers=Allprocedures conductedin Class II i BSCs orClass I or II BSCs incombination with full-body, air-supplied,posit ive pressurepersonnel suit

BSL-1 plus: autoclaveavailable

BSL-2 plus:9 Physical separat ionfrom access corr idors9 Self-closing, doubledoor access9 Exhausted air notrecirculated9 Negat ive air f low intolaboratoryBSL-3 plus:9 Separate building orisolated zone9 Dedicated supply /exhaust, vacuum, anddecon systems9 Other requirementsas out l ined in therecommended text

Based on 'Biosafety in Microbiological and Biomedical Laborator/es' with kind permission of the Editor, JW Richmond

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appendix a - biosafety in the virology laboratory

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