Unit 1-Lecture 2 Basic Laboratory Safety: Contemporary Clinical Immunology and Serology

Contemporary Clinical
Immunology and Serology
CHAPTER 22
Unit 1-Lecture 2
Basic Laboratory
Safety
Contemporary Clinical Immunology and Serology

Kate Rittenhouse-Olson | Ernesto De Nardin
Basic Laboratory Safety
• Safety equipment
• Personal protection equipment (PPE)
• Safety procedures
• Safety and the law
• Needlestick injuries
• Postexposure treatment
• Chemical and radioactive hazards
• General safety
Common Laboratory Hazards
• Contamination
 Biological
 Radioactive
• Burns
 Chemical
 Heat sources

Common Laboratory Hazards
• Sharps
 Needlesticks
• Other physical trauma

Examples of Safety Equipment
• Warning labels
• Containment hoods
• Emergency showers
• Eye washes
• First-aid kits
• Waste-disposal containers
• PPE

Warning Labels
• Identifies potential hazards

• Posted on
 Laboratory doors
 Laboratory equipment
 Containers of chemicals
 Biological samples
 Radioactive materials

National Fire Protection
Association (NFPA) Diamond
• Health risks (blue)
• Flammability risks
(red)
• Reactivity risks
(yellow)
• Special risks
(white)

National Fire Protection
Association (NFPA) Diamond
• Numbers in each color indicate level of
risk
 0 in red indicates nonflammable
reagents
 4 in red indicates dispersal in air, burns
at all temps
• Contact information

Chemical Containment Hoods
• "Fume hoods"
• OSHA
 Device enclosed on five sides
• Partial covering or sash
• Work is done within the hood
• Technician's arms/hands only inside
hood

• Negative pressure
 Air drawn in from lab, exhausts into
separate air-handling system
 Individuals not exposed to fumes from
reagents
• National Research Council
 People working together in fume hood
• 2.5 linear feet of space

• Ventilate 60–100 linear feet per minute

 Not too high ( turbulence)
• Lab hood as work space, not storage
space
 Clutter causes accidents and increases
chance of fire when working with
flammable reagents

Figure 22.2
Typical laboratory chemical hood.

Biological Containment Hoods
• Protects the operator

• Avoids contaminating sample/specimen
• Air through HEPA filter (0.3 μm)

Laminar Flow Hood
• Forced air constantly runs downward at

the opening of the hood
 Prevents entry or exit of contaminating
particles
• Hoods can be designed just to protect
sample from outside contamination and
may not protect room from the sample

Laminar Flow Hood
• Germicidal ultraviolet lights inside hood

 Maintain sterile conditions
 Turned off when specimen is handled
• Biological safety cabinet
 Appropriate air flow
• Position—no drafts from doors, windows,
or traffic

Figure 22.3
Laminar flow hood.

Emergency Showers and
Eye Wash Stations
• Operator contamination
 Large spills on lab worker
 Clothing set on fire by flammable
chemicals
 Splashes in eyes

Emergency Showers and
Eye Wash Stations
• Equipment location
 Unrestricted pathway
• Urgent need to shower or eye wash
• Less than 10 seconds to reach, if strong
acids or bases are used
• Less than 100 feet from injury

Figure 22.4
Typical laboratory emergency shower.

Eye Wash Stations
• Design
 Able to operate with impaired vision
 Large handles
 Provide copious amounts of water
 Directed to face, eyes

Eye Wash Stations
• Two types
 Plumbing permanently connected to wall
 Dispense fluid by gravity flow
• Devices contain water or saline

Figure 22.5
Emergency eyewash station.

Safety Containers
• Safe and proper disposal

 Biological hazards
 Hypodermic needles, syringes
 Glass and sharps
• Do not discard in conventional trash
• Collect and process according to
regulations

Sharps Safety Containers
• Puncture proof that can be autoclaved

• Openings that do not allow retrieval
once items are disposed
• No overfill
• Other sharps (broken glass, surgical
blades, glass slides)
 Dispose in puncture-proof containers
 Containers can also be autoclaved

Figure 22.6
Typical disposal container for sharps such as needles.

Figure 22.7
Typical container for the disposal of broken glass or other sharps.

Safety Containers
• Disposing biological materials

 Leakproof, autoclavable plastic bag
 Clearly labeled with biohazard warning
 Once autoclaved dispose in labeled
biohazard disposal boxes

Figure 22.8
Typical box used for the disposal of bagged, sealed, and autoclaved biological waste.

Personal Protection Equipment
• Goggles
• Face shields
• Lab coats
• Lab gowns
• Face masks
• Gloves

Figure 22.9
Operator using personal protection equipment.

Laboratory Coats and Gowns
• Disposable or multiple use

• Prevent biological contamination (body
fluids [blood, urine, saliva, excretions])
to operator and clothing
• Fully buttoned
• Preferably fluid resistant

Laboratory Coats and Gowns
• Tight sleeves
 Fit beneath the gloves
 Removed and changed patient to patient
(if patient contact is required)
• Wash frequently if multiple use
• Single use
 Discard immediately after use

Gloves
• Different types and sizes

 Sterile surgical gloves
 Nonsurgical latex gloves
 Simple vinyl gloves
• Nonsterile gloves are sufficient for most
labs
• Protection from blood, body fluids,
excreta, other potentially contaminated
items
Gloves
• Gloves alone are not sufficient for

protection
• Wash hands thoroughly both before
and after wearing gloves
• Change gloves from patient to patient
• Remove immediately after use and
dispose

Gloves
• Snug, well-fitting
• Extend over cuffs or sleeves of lab
coats and gowns to prevent
contamination
• Never kept on when leaving lab or
when changing from one station of lab
to another

Gloves
• Never worn while:

 Opening doors
 Touching environmental surfaces
 Operating lab equipment away from
operating station

Latex Sensitivity
• Latex
 Most common glove material
 Allergy
• Type 1 hypersensitivity most common
• Reactions—red, itchy skin, hives,
coughing, wheezing

Latex Sensitivity
• Latex
 Allergy
• Avoid using in areas where gloves are
changed
• Type IV hypersensitivity reactions
(contact dermatitis)

Masks, Goggles, and Face Shields
• Protects operator from splashes

 Biological samples
 Potentially airborne aerosols and
pathogens

Masks, Goggles, and Face Shields
• Worn when manipulating biological

samples
 Prevent contamination of mucosa
(mouth, nose, eyes)
• Alternative to goggles or face shields
 Lab bench shield

Personal Care and Behavior
• Tie back long hair

 Reduces risk of contamination
 Reduces risk of entrapment in moving
equipment
• Closed-toe shoes at all times
• Nothing placed in mouth while in lab

Vaccination
• OSHA 1991
 Hepatitis B vaccine
• Employers must offer without charge to
those at risk
 Rubella, mumps, measles, influenza
• Recommends vaccinate hospital
employees
• Especially women of childbearing years

Universal Precautions
• Assume all patients and samples are

potentially infectious and possible
carriers

• Guidelines
 Wear PPE (gloves, masks, coats, or
gowns)
• When collecting blood or handling body
fluids that may be contaminated with
blood
• When there is a possibility of blood
splashing on mucosal surfaces

• Guidelines extended to other body

fluids
 Semen
 Vaginal secretions
 Synovial fluid
 Amniotic fluid

• Guidelines extended to other body

fluids
 Cerebrospinal fluid
 Pleural fluid
 Peritoneal fluid
 Pericardial fluid

Universal Precautions Guidelines
• Wash hands after removing gloves

• Never pipette by mouth
• Transport sample in leakproof container
with lid
• Decontaminate work surfaces after
working with blood or body fluid

Universal Precautions Guidelines
• Disinfect biological spills with a fresh

solution of 1:10 dilution bleach or with
commercial disinfectant
• Decontaminate material before disposal
• Do not wear protective clothing outside
lab

Hand Washing
• Basic yet most effective safety

procedure

Hand Washing
• APIC and CDC guidelines

1. Wet hands with warm water
2. Apply soap and spread over hands
3. Rub hands together for 20 seconds,
hard enough to generate friction; rub
all surfaces and under fingernails
• Wash from wrist area to tips of fingers

Hand Washing
• APIC and CDC guidelines

4. Rinse thoroughly
• Water flowing in downward direction
(wrists to fingers), carrying
contaminated water away from operator
and into sink
5. Dry with towels
6. Turn off sink with a towel

Hand Washing
• Wash any time contamination is

suspected
• Wash before and after wearing gloves
• Wash between patients and between
different samples

Hand Washing
• Wash when gloves are changed

• Wash before leaving lab, before food
and drink handling, and after using
restrooms

Sample Handling
• Assume every sample is source of

contamination
• Spread of infection follows a specific
pattern
 Source
 Method of transmission
 Host

Sample Handling
• Most common source of infection in

clinical lab
 Biological specimens

OSHA Directives for Laboratory
Behavior
• No mouth pipetting
• No eating, drinking, smoking
• No applying of cosmetics or lip balm
• No handling contact lenses in lab

OSHA Directives for Laboratory
Behavior
• No storage of food or drink in lab
refrigerators or lab bench tops
• Chipped or broken glassware should be
discarded

Safety and the Law
• OSHA
 Occupational Exposure to Bloodborne
Pathogen (BBP) Standard (29 CFR
1910.1930)
• Regulations to instruct and protect health
workers about exposure to BBPs
• Develop and implement exposure control
plan

Safety and the Law
• OSHA
1910.1930)
• Provide health workers with free lab coats,
gowns, and other personal safety
equipment

Safety and the Law
• OSHA
1910.1930)
• General lab safety equipment (warning
labels)

29 CFR 1910.1930 Regulations
• No eating and drinking in lab

• Immunization against hep B must be
available, no charge
• Laundering services must be provided
free for multiple-use items (coats,
other protective clothing)
• Maintain records of health workers
training

29 CFR 1910.1930 Regulations
• Establish well-defined procedures and

follow-up should worker be exposed to
BBP
 Pathogenic microorganisms present in
human blood can cause disease in
humans, not limited to
• Hepatitis B virus (HBV)
• Human immunodeficiency virus (HIV)

Needlestick Injury Protocols
• Major pathogens of concern

 HIV
 Hepatitis B
• Vaccination reduces risk 90–95%
• Survives on surfaces > 1 week
• Contamination by inanimate object can
occur

Needlestick Injury Protocols
• Major pathogens of concern

 Hepatitis C
• Risk 0–7%
 Hepatitis D
• Minor concern
 Tetanus
• http://www.ccohs.ca/oshanswers/disea
ses/needlestick_injuries.html

Transmission Rate
• Rate of occupational transmission

 HIV
• 0.3% needlestick
• 0.09% mucous membrane exposure
 Hepatitis B
• 6–24% if nonimmunized
 Hepatitis C
• 1–10%

Treatment
• Needlestick
 Squeeze one drop blood
 Wash hands
 Apply alcohol wipe
 Stop
• No harsh manner; don't want white cells
to area

Treatment
• Mucous membrane
 Wash/flush with large amounts of saline

Report
• Report to supervisor
• Go to employee clinic
 Need history to determine risk
• Immunization record (hepatitis and
tetanus)
• Previous occupational exposure to body
fluids
• IV drug use
• Sexual history

Report
• Go to employee clinic
 Need history to determine risk
• Body piercing or tattooing
• Receiving blood or blood products
• Dialysis history
• Travel outside U.S. within last year
 Physical for baseline lung, liver, lymph
node status

Risk Factors
• Failure to follow Universal precautions

• Use of equipment without safety
feature
• Performing exposure-prone tasks

Laboratory Testing
• Source patient with consent

 HIV
 Hepatitis B
 Hepatitis C
 AST (aspartate aminotransferase)
 ALT (alanine aminotransferase)
 Alkaline phosphatase

Laboratory Testing
• Health-care worker
 Hepatitis B surface antibody
• Did vaccine work?
 HIV
 Hepatitis C antibody testing
• Baseline
• 2 weeks
• 4 weeks
• 8 weeks
Laboratory Testing
• Prior to initiating retroviral therapy

 Stat pregnancy test
 Liver function
• AST/ALT, alkaline phosphatase, bilirubin

Laboratory Testing
• Prior to initiating retroviral therapy

 Kidney function
• Serum creatinine/BUN levels, urinalysis
with microscopic analysis
 General health
• CBC with diff and platelets

Immediate Care
• Irrigate and clean wound

• Tetanus shot
• Hepatitis B prophylaxis if not
immunized
• Hepatitis A prophylaxis (rarely needed)
• HIV risk assessment

Figure 22.10
Different exposure codes for potential HIV infection.

Figure 22.11
Risk assessment codes for an HIV-infected patient.

Postexposure Prophylaxis (PEP)
• Basic regime four weeks

 Zidovudine (AZT) and lamivudine (3TC),
a nucleoside analog reverse
transcriptase inhibitor

Postexposure Prophylaxis (PEP)
• Expanded regime
 Zidovudine (AZT) and lamivudine (3TC)
 Indinavir (protease inhibitor)
 Nelfinavir (protease inhibitor)
• Interferon ribavirin
 Decreases risk by 40%

After Postexposure Prophylaxis
(PEP)
• Abstain from sex
 6 months or until third negative test
• Breast feeding discouraged
• Do not donate blood
• Receive follow-up care for 6 months to
determine seroconversion status

Needlestick with Hepatitis C risk
• 1.8% chance of seroconverting if

needlestick
• Splash to eyes or mouth risk very low
• Test both for anti-HCV and ALT
• Follow 4–6 months to look for antibody

Needlestick with Hepatitis C risk
• Follow 4–6 weeks if testing for HCV

RNA
• Run confirmatory test
• http://www.cdc.gov/hepatitis/HCV/HCV
faq.htm

Prevent Exposures
• Use safer techniques

• Do not recap needles
• Dispose used needles in sharps
containers
• Use devices with safety features
• Wear gloves
• Wear eye and face protection
• Wear gowns

Other Diseases Spread (Rarely) by
Needlestick
• Blastomycosis
• Brucellosis
• Cryptococcosis
• Diptheria
• Cutaneous gonorrhea
• Herpes
• Malaria
• Mycobacteriosis
Other Diseases Spread (Rarely) by
Needlestick
• Mycoplasma caviae
• Rocky mountain spotted fever
• Sporotrichosis
• S. aureus
• S. Pyogenes
• Syphilis
• Toxoplasmosis
• Tuberculosis
Chemical Hazards
• PPE (laboratory coats, masks, face

shields, gloves) to avoid contact with
skin, mucous surfaces, and clothing
• Handle flammable or toxic vapor
chemicals inside fume hood
• Read labels on reagent bottles
 Learn potential hazards and proper
handling instructions
• Material Safety Data Sheet (MSDS)
Material Safety Data Sheet
(MSDS)
• Provided by manufacturer
• Chemical properties

(MSDS)
• Physical properties
 Melting point
 Boiling point
 Flash point
 Reactivity
 Toxicity
 Health effects

(MSDS)
• Other info
 First-aid procedures
 Specific protective equipment
 Storage and disposal
 How to handle spills

Chemical Spills
• Wash affected area immediately

 Copious amounts of water for 15–20
minutes
• First aid should follow

Chemical Spills
• Be aware of shower/eye-washing
station locations
 Practice reaching them quickly, even
under impaired conditions
• Use appropriate "spill kit" to clean spills
 MSDS has what absorbent or
neutralizing material to use

Liquid Nitrogen
• Used to preserve cells and tissues

• If spilled on skin, can cause cold-
contact burns

Liquid Nitrogen
• Liquid nitrogen
 Room temperature—quickly becomes
gas
• Liquid to gas volume ratio is 1:694
 Replaces oxygen in room; acts as
asphyxiant
 Dangerous when room oxygen levels
< 16%
 Store with oxygen sensor

Radioactive Hazards
• Radioactivity in routine clinical lab

assays very low
• Effect of radiation is cumulative
• Avoid or minimize exposure
• Nuclear Regulatory Commission (NRC)
regulates radioactive materials

Nuclear Regulatory Commission
(NRC)
• Dictates amount of radiation allowed in
lab
• Regulates different isotopes used for
assays
• Regulates monitoring, tracking, and
disposal of radioactive materials

NRC Regulations
• Labs are required to keep records of

 Quantity of materials used
 Type of isotopes used
 Disposal of waste
 Purchase of materials
 Disposal of unused materials
• Records are kept in a book or binder
provided by the radiation protection
services of the lab or hospital
Figure 22.12
Radioactive protection services record book and two badge-style dosimeters.

NRC Regulations
• PPE (gloves, masks, and gowns) must

be used
• Dosimeters
 Devices that record amount of
radioactivity
 Assigned to single individual user
 Worn as badges or rings
 Periodically monitored by radiation
protection services
NRC Regulations
• Work is carried out in designated areas

• Areas are labeled for use of radioactive
materials
 Monitored routinely (1x month) for
contamination with periodic swab tests

NRC Regulations
• Results recorded in radiation-protecting

services record book
• Disposal is carefully regulated and
monitored
 Both local regulation and NRC
• Amount of radioactivity and
radioisotopes involved varies according
to type of material (solid, liquid,
volatile chemical)
General Safety
• Workers are trained in how to use

safety equipment
• Know the location of fire blankets, fire
extinguishers, showers, eye wash
stations, decontaminating solutions,
spill kits, and first-aid kits

General Safety
• Understand fire evacuation route

• Designate an evacuation meeting place
 "Role call" (ascertain if anyone unable to
leave building)
• Electrical equipment should not be
plugged into extension cords or into
"gang" plugs

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Contemporary Clinical

Immunology and Serology

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

• Identifies potential hazards

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

• Ventilate 60–100 linear feet per minute

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

• Protects the operator

Contemporary Clinical Immunology and Serology

• Forced air constantly runs downward at

Contemporary Clinical Immunology and Serology

• Germicidal ultraviolet lights inside hood

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

• Safe and proper disposal

Contemporary Clinical Immunology and Serology

• Puncture proof that can be autoclaved

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

• Disposing biological materials

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

• Disposable or multiple use

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

• Different types and sizes

• Gloves alone are not sufficient for

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

• Never worn while:

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

• Protects operator from splashes

Contemporary Clinical Immunology and Serology

• Worn when manipulating biological

Contemporary Clinical Immunology and Serology

• Tie back long hair

Contemporary Clinical Immunology and Serology

Contemporary Clinical Immunology and Serology

• Assume all patients and samples are

Contemporary Clinical Immunology and Serology