Bloodborne Pathogens and Biosafety powerpoint
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Transcript of Bloodborne Pathogens and Biosafety powerpoint
USC Environmental Health & Safety
GoalSafety: 385,000 percutaneous injuries/yr in hospitals1990 OSHA estimate: 9,000 bloodborne infections/yr, 200
deathsLaboratory acquired infections - Sulkin and Pike: 3,921
cumulative cases, including brucellosis, typhoid, tularemia, tuberculosis, hepatitis, and venezuelan equine encephalitisLess than 20% associated with known accidentLab environment more hazardous than nature due to
culturing, propagation of infectious agentsRecent events at U Chicago, Texas A&MCompliance with Safety Regulations: Cal-OSHA BBP
Standard, 1992Education, prevention
Lab acquired vaccinia infection, Virginia 2008
Worked in cancer research labHandled mice infected with vaccinia virusNot vaccinated
What are Bloodborne Pathogens?microorganisms ( such as viruses or bacteria)
that are carried in blood and can cause disease in people
malaria, syphilis, Hepatitis B Virus(HBV), Hepatitis C Virus, Human Immunodeficiency Virus (HIV)
Am I at risk?Work with blood or OPIMOPIM - semen, vaginal secretions, saliva in
dental procedures, CSF, or other internal body fluidsOrgans, unfixed human tissueHBV, HCV, HIV tissue culturesHuman cell lines
Human Cell LinesCell lines may be infected or become
infected/contaminated in subsequent handling/passaging
LCMV- Researchers infected working with nude mice (infected tumor cell line)
“We recommend that all human cell lines be accorded the same level of biosafety consideration as a line known to carry HIV… Thus, it is best to use caution when handling any human cell line”
Universal Precaution
Low Risk MaterialsUrine, feces, vomit, tears, sweat, sputum,
nasal secretionsUnless visibly contaminated with blood
Routes Of TransmissionSexual contactSharing needlesBlood Transfusion (very rarely in countries where
blood is screened)Birth: Expose through amniotic fluid, blood during the
birthing process, and (less likely) through breast milk Accidental puncture from contaminated
needles, glass or other sharpsContact between broken or damaged skin and
infected body fluidsContact between mucous membranes and infected
body fluids
Common procedures where exposure may occurHandling containers of blood, fluid, tissue,
or culturesPhlebotomyPipetting, mixing, or handling blood, fluid, or
tissueCleaning blood/body fluid spillsHandling contaminated sharps or other
contaminated wastePuncture from improperly disposed needle
Injections/inoculations~89% occupationally acquired HIV
Determinants of Disease TransmissionIndividual receiving the doseVirulence / type of pathogenSize of delivered dose (concentration)Route of exposureType of bodily fluid Whether post exposure prophylaxis was
administered
Diseases caused by BBPHepatitis BHepatitis CAIDS(HIV)
Hepatitis BInflammation of the liverRisk of HBV infection after a single positive needle stick is 30%Durable virus (7 days outside body)Acute or chronicMore than 350 million people are chronic carriers of HBV
worldwide (CDC) 800,000–1.4 million persons in the United States
have chronic HBV infection
Annual number of unreported infections may be 10 times greater than the number of reported (many HBV infections are either asymptomatic or never reported)
rate of new HBV infections has declined ~80% since 1991
Hepatitis BSymptoms (weeks-~6months) – Fever,
Jaundice, fatigue, abdominal pain, loss of appetite, nausea, vomiting, joint pain, dark urine
Complications - Cirrhosis (scarring) of the liver, liver cancer, liver failure, and death
Prevention - Hepatitis B Vaccine, PEP (Plasma fractionation of Hep B Antibody ), Universal Precautions
http://phil.cdc.gov/phil/home.asp
HEP B VaccineUSC offers the vaccines to employees
FREE OF CHARGESeries of 3 inoculations (shots) and Titer
checkAfter vaccination cellular immunity
persists (despite low antibody levels)
Hepatitis CAfter a needlestick or sharps exposure to
HCV-positive blood, the risk of HCV infection is approximately 1.8%
75-80% chronic 3.2 million persons in the U.S.- chronic HCV
infectionVaccine? NoPEP? No
Hepatitis C VirusOf every 100 persons infected with HCV,
approximately 75–85 will go on to develop chronic infection60–70 will go on to develop chronic liver disease5–20 will go on to develop cirrhosis over a period of 20–30 years 1–5 will die from the consequences of chronic infection (liver cancer or cirrhosis)
8,000–10,000 deaths each year in the United States (chronic)
Hepatitis C Symptoms - Jaundice, fatigue, abdominal
pain, loss of appetite, nausea, dark urineComplications – Chronic liver disease, deathTransmission: Occurs when blood or body
fluids from an infected person enters the body of a person who is not infected
Prevention – Universal precautions
HIVAttacks Immune SystemInfects/Kills T-Helper Cells (CD4)Final Stage of HIV Infection: AIDS – Acquired
Immunodeficiency SyndromeOpportunistic infections – Kaposi’s sarcoma,
CMV Workers have been infected with HIV after
being stuck with needles containing HIV-infected blood or after infected blood gets into a worker’s open cut or a mucous membrane
Electron microscope image of HIV, seen as small spheres on the surface of white blood cells.
http://www.biologyimagelibrary.com/imagelibrary/images/12646_1_PHIL_1843_lores_BIL260805.JPG
HIVFragile retrovirusRisk of transmission after percutaneous exposure to
HIV infected blood is approximately .3% ROT after splash to eye/mouth/nose ~.1%57 healthcare workers with documented
occupationally acquired HIV infectionCDC ~56,300 new HIV infections in the
US in 2006
http://www.agen.ufl.edu/~chyn/age2062/lect/lect_14/Lect_14.htm
Healthcare Personnel with Documented and Possible Occupationally Acquired AIDS/HIV Infection, by Occupation, 1981-2006
Occupation Documented Possible
Nurse 24 35
Clinical Laboratory Worker
16 17
Nonsurgical Physician
6 12
Non-Clinical Laboratory Technician
3 -
Housekeeping or Maintenance
2 13
Dentist or Dental Worker
- 6
U.S. Infect Control Hosp Epidemiol 2003;24:86-96.
What is risk with lentiviral vectors?Potential for generation of replication-
competent lentivirus (RCL)Potential for insertional
mutagenesis/oncogenesisHost rangeMitigated by the nature of the vector system
(and its safety features) or exacerbated by the nature of the transgene insert encoded by the vector.
Universal PrecautionTreat all blood and body fluids as if infectious For labs: Biosafety Level 2 (BSL2)
containmentDefined by CDC/NIHCombination of laboratory practices and
techniques, safety equipment, and laboratory facilities
Many BSL2 safety procedures extend to non-lab settings
BSL2: Standard Microbiological PracticesAccess to lab limited during experimentsWashing hands after handling viable materials,
after removing gloves, and before leaving labNo eating, drinking, smoking, handling contacts,
or applying cosmeticsSafe handling of sharps, plastic substituted
for glassMinimize splash/aerosolsDecontaminate work surfaces daily and after spillsProper disposal of contaminated items as
biohazardous waste
Biohazard Waste DisposalGauzePlastic serological pipettesPlastic pipette tipsPetri dishesCulture vials/platesCounter top covers Contaminated PPE
SharpsContaminated broken glasswareScalpels, blades, capillary tubesSlides & cover slipsGlass pipettesGlass capillary tubesNeedles and syringes
Liquid WasteDecon. tissue and media with 10% bleach
solution~ 15 minute contactMay then be poured down drain USC does not have a permit to autoclave
medical waste!
BSL2: Special PracticesOnly those who have been advised of potential hazards may
enter labBiohazard sign when agent in use, include relevant contact
information, info on agent, what to do if exposed, PPEPI ensures all are appropriately trained on potential hazards of
work involved, how to prevent exposure, offered appropriate immunizations
Training on hazards, exposure prevention, SOP’sSharps precautions, engineered sharps, handling broken
glasswareInfectious agents placed in container with cover that prevents
leakage during collection, handling, processing, storage, transport, shipping
Decontamination of equipment/work surfaces Report exposures to PI, seek treatment
Safety engineered sharps2001 Needlestick Prevention Act
LabelingInternational biohazard symbolBiohazardous wasteDoor signsIncubatorsFreezersRefrigeratorsShipping boxes
DecontaminationWork surfaces, tools, and equipment1:10 bleach solutionLysol or other EPA registered disinfectant15 minute minimum decontamination time
Handling, storage, transport proceduresSpecimens must be placed in a container which
prevents leakage during collection, handling, processing, storage, transport, or shipping (use secondary container), container must be labeled with biohazard symbol
BSL2: Safety EquipmentRequires the use of a BSC for large volumes or potential
aerosol generationUse of centrifuge safety cups/sealed rotors for aerosol
containmentFace protection for anticipated splashes or sprays when
outside BSCLab coats, leave before leaving labGloves whenever potential contact with hazardous agents
Biosafety Cabinets
Biosafety CabinetsSweeping motions of armsDisinfect surfaces before and after useFront intakeUVBunsen burners alternatives (i.e. sterile
loops, touch-plate microburners)Substitute plasticware for glassware or
other sharp instruments whenever possible (i.e. pasteur pipettes)
Must be professionally decontaminated/recertified before moving
Recertify annually
BSL2: Lab facilities
Lockable doors Sink for handwashing No carpets/rugs Chairs covered in non-porous material BSC away from doors Eyewash station
Emergency procedures for spills Inform others in lab and restrict accessLeave area and allow aerosols to settleDon personal protective equipment If broken glass present, never handle directly by hand, use
forceps to remove and dispose into sharps containersPlace paper towels/absorbent over spillApply disinfectant(10% bleach) directly onto paper towels
and allow 15 minute contact time to disinfectClean spill by wiping around the perimeter and moving
inwardsDiscard materials as biohazardous wasteWash hands with soap and water
Emergency procedures for exposuresFlush affected area with water for 15 minutesInform supervisorKnow where to get treatment
Avoid working alone in lab!
Treatment LocationsHSC
Business HoursInternal Medicine (HCC)
1520 San Pablo St.Los Angeles, CA 90033
(323) 442-5100
After HoursWhite Memorial Hospital
1904 Bailey Street, Suite 100Los Angeles, CA 90033
(323) 222-9675
Exposure IncidentReport incident to supervisor immediately and
seek medical treatment! File Supervisor’s report of injuryFile Sharps Injury FormReport all exposure incidents to IBC
USC Exposure Control Plan
http://capsnet.usc.edu/LabSafety/BioSafety/BloodBornePathogensProgram
How to elim/minim exposure to human/NHP materials that might contain BBP
Outlines PI responsibility: Update ECP, GLS/BBP, Hep B vaccination
Methods of Compliance: Safety EquipmentGood Work Practices : PPE, hand washing,
labelingMedical Surveillance Program: PE eval./follow up
Contact InformationEnvironmental Health and Safety
323.442.2200(office)[email protected], CHP 148