Sikkerhedsforanstaltninger og driftsprocedurer i et (A) BSL-4 Laboratorieundersøgelser: 4. Medicinsk Imaging Procedurer
Summary
Here, we present an overview of the preparation and animal handling procedures required to safely perform medical imaging in an animal biosafety level 4 laboratory. Computed tomography of a mock-infected guinea pig illustrates these procedures that may be used to evaluate the disease caused by a high consequence pathogen.
Abstract
Medical imaging using animal models for human diseases has been utilized for decades; however, until recently, medical imaging of diseases induced by high-consequence pathogens has not been possible. In 2014, the National Institutes of Health, National Institute of Allergy and Infectious Diseases, Integrated Research Facility at Fort Detrick opened an Animal Biosafety Level 4 (ABSL-4) facility to assess the clinical course and pathology of infectious diseases in experimentally infected animals. Multiple imaging modalities including computed tomography (CT), magnetic resonance imaging, positron emission tomography, and single photon emission computed tomography are available to researchers for these evaluations. The focus of this article is to describe the workflow for safely obtaining a CT image of a live guinea pig in an ABSL-4 facility. These procedures include animal handling, anesthesia, and preparing and monitoring the animal until recovery from sedation. We will also discuss preparing the imaging equipment, performing quality checks, communication methods from “hot side” (containing pathogens) to “cold side,” and moving the animal from the holding room to the imaging suite.
Introduction
Missionen for National Institute of Allergy og smitsomme sygdomme (NIAID) Integreret Research Facility ved Fort Detrick i Frederick MD (IRF-Frederick) er at udføre nye smitsomme sygdomme forskning for at forstå de kliniske sygdomsprocesser, der korrelerer med sværhedsgraden af mikrobielle-induceret sygdom. IRF-Frederik har en unik evne til at udføre medicinsk billedbehandling i dyremodeller af høj konsekvens patogener i en ABSL-4 laboratorium 1. De billeddannende modaliteter til rådighed for forskere omfatter: computertomografi (CT), magnetisk resonans billeddannelse (MRI), positron emission tomografi (PET), enkelt foton computertomografi (SPECT), ultralyd, røntgen, og fluoroskopi. Forskere bruger tilgængelige billeddiagnostiske kapacitet til at overvåge sygdomsprogression og evaluere effekten af interventioner, såsom narkotika behandling og vaccination, i longitudinelle studier.
De billeddiagnostiske metoder på IRF-Frederick var specifiktdesignet til at holde de centrale dele af udstyret uden for høj inddæmning 2,3 og tilgængelige for vedligeholdelse og reparation. Dette design adskiller imaging suite til "hot" (indeholdende patogen) og "kolde side". For at opnå denne adskillelse blev specielt konstruerede rør konstrueret til at udvide høj indeslutning plads i boringerne af hver afbildningsmodalitet (figur 1). Ud over at give biologisk indeslutning, disse rør beskytter billedudstyr fra gasser og kemikalier, der anvendes til at rense den høje inddæmning laboratorium. Imaging forskere og teknologer betjene scannere fra "kolde side", mens Comparative Medicine (CM) personale håndtag og overvåge dyr på den "varme side". Da CM personale skal arbejde tæt sammen med imaging forskere til at koordinere disse eksperimenter, kan denne adskillelse resultere i kommunikations udfordringer.
Efter at have vurderet muligheder, CM personale var udeudstyret med Bluetooth øre stykker, der sender korte bølgelængde ultra-højfrekvente radiobølger til telefoner, der anvendes til at kalde den billeddannende personale uden for inddæmning. På grund af udformningen af anlægget, skulle installeres i hvert af værelserne til at overvinde interferens forårsaget af de lag af cement og stål mellem "hot" og "kolde sider" trådløse adgangspunkter. Således kommunikation mellem CM personale iført støjende positiv-trykdragter og billedbehandling personale uden høj indeslutning er nu pålidelige. Kameraer er også blevet installeret på den varme side af de billeddannende værelser til afbildning personale til at se aktiviteten på den "varme side". Med kameraerne, kan den billeddannende personale guide CM teknikere med animalske positionering eller eventuelle ændringer i sidste øjeblik til det billeddannende protokollen.
Alt arbejde i IRF-Frederick ABSL-4 suit laboratorium kræver medarbejdere til at bære positivt tryk indkapsler jakkesæt 4. Iført disse dragter reducerer mobiliteten, og den tunge sentx handsker knyttet til kulør plus op til tre yderligere lag af handsker kompromiser fingerfærdighed. Resultatet er, at procedurerne tager længere tid at gennemføre og opgaver, der kræver finmotorisk færdighed er meget vanskeligere. Som biosikkerhed niveau stiger, håndtering af dyr og manipulationer bliver mere udfordrende og tidskrævende, især med små dyr. Procedurer i en ABSL-4 laboratorium kan tage op til 2-3 gange længere end en ABSL-2 laboratorium.
Formålet med denne artikel er at visuelt demonstrere de udfordringer forbundet med billedbehandling dyremodeller i en ABSL-4 miljøet ved hjælp CT-scanning procedure af et marsvin som et eksempel.
Protocol
Representative Results
Discussion
Tidligere artikler i denne serie har understreget omfattende uddannelse, sans for detaljer, sikkerhedsprocedurer, og yderligere teknisk kontrol, der kræves for at arbejde sikkert i en maksimal inddæmning laboratorium 12,13. Udførelse arbejde sikkert er den højeste prioritet i disse laboratorier. Denne filosofi er endnu vigtigere, når man arbejder med levende dyr på grund af yderligere farer såsom muligheden for smittede dyr at påføre bid eller ridser eller aerosoldannelse 7. Disse procedur…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
The content of this publication does not necessarily reflect the views or policies of the US Department of Health and Human Services (DHHS) or of the institutions and companies affiliated with the authors. This work was funded in part through Battelle Memorial Institute’s prime contract with the US National Institute of Allergy and Infectious Diseases (NIAID) under Contract No. HHSN272200700016I. M.R.H., K.J., D.P., L.B., and J.W. performed this work as employees of Battelle Memorial Institute. Subcontractors to Battelle Memorial Institute who performed this work are: R.B., an employee of Charles River Laboratories – Insourcing Solutions; L.K. and M.R.L., employees of MEDRelief Staffing Inc.; M.G.L. as an employee of Lovelace Respiratory Research Institute, Inc.; and J.H.K. as an employee of Tunnell Government Services, Inc.
Materials
| Micro-Chem Plus | National Chemical Laboratories | 255 | |
| CT scanner | Philips Healthcare | ||
| CT phantom | Philips Healthcare | ||
| Isovue-300 (CT contrast reagent) | Bracco Diagnostics | NDC 0270-1315-30 | |
| Ventilated rack | Lab Products | ||
| Micro-isolator cage | Lab Products | ||
| Biosafety cabinet | Nuaire | ||
| Anesthesia machine | SurgiVet | WWV9000 | |
| Anesthesia induction box | VetEquip | ||
| Anesthesia mask | Henry Schein | ||
| Isoflurane | Henry Schein | ||
| Waste gas scavenging canister | Fisher | F/AIR | |
| Holding cushion | |||
| Ophthalmic ointment | |||
| Vital signs monitor | Bionet | BM3Vet | |
| Mobile phone | Spectralink | 8440 | |
| Blue Tooth ear piece | |||
| Wireless access points | |||
| Sperian positive-pressure suit | Honeywell Safety Products | BSL 4-2 | |
| Outer suit gloves (latex, Ansell Canners and Handlers) | Fisher | 19-019-601 | |
| Outer suit gloves (nitrile/rubber, MAPA) | Fisher | 2MYU1 | |
| Scrubs | Cintas | 60975/60976 | |
| Socks | Cintas | 944 | |
| Duct tape | Pack-N-Tape | 51131069695 | |
| Towels | Cintas | 2720 | |
| Zip lube | Amazon | B000GKBEJA |
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