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Disinfection

disinfection and steralization in clinical lab

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8 views29 pages

Disinfection

disinfection and steralization in clinical lab

Uploaded by

dina lithy
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Disinfection & Sterilization

Dr. : Dina Ramadan


Microbiologist In Central Health Laboratory
Ministry of Health
Quality Manager
Transmission Risk
# of virus particles/ml of blood Risk of infection, due to injected
Virus exposure
6 13
Hepatitis B 10 -10 1 in 3 (30%)
(Millions – trillions)

6
Hepatitis C 10 1 in 30 (3%)
(Millions)

3
HIV (AIDS) 10-10 1 in 300 (0.3%)
(Tens – thousands)

Hepatitis A virus it can survive outside the body for months.


Hepatitis B virus can survive outside the body at least 7 days.
HCV, can live outside the body for up to 4 days. However, many experts think it
usually survives up to 16 hours at room temperature.
HIV may persist for at least a week in dried blood at 4°C, it survive for up to four
weeks in syringes ,and may survive in dried blood at room temperature for up to
five or six days
Definitions
Sanitization:
Scrubbing to remove blood, mucus, or
media where pathogens can grow

Disinfection:
It is a procedure intended to reduce
microorganisms as far as possible (but not
bacterial spores) by physical and chemical
means

Sterilization:
the complete removal or destruction of all
forms of microbial life including bacterial
spores
Remember :
No absolute assurance that there is 0 microorganism
Sanitization
• First step in disinfection and sterilization
for other equipment
• Scrubbing with a brush and detergent to
remove blood, and other contaminants or
media where pathogens can grow
Chemical Disinfectant
Alcohols

• There are two water-soluble alcohol can be used in


lab: ethyl alcohol and isopropyl alcohol.
• Concentration required : 60-90% in water
• Mode of its action : it denaturing bacterial proteins
• its Flammable, Can be removed from disinfected
articles by flaming.
Alcohols effect on bacteria

• they are bactericidal against for the vegetative


forms of bacteria (Gram + and Gram -);
• also they are tuberculocidal, fungicidal, and
virucidal against enveloped viruses (HIV, influenza
,herpes viruses).

• not effective against bacterial spores and have


limited effectiveness against nonenveloped viruses
( adenovirus, rotavirus)
Alcohols
• Higher concentrations are
less effective as the action
of denaturing proteins is
inhibited without the
presence of water
Hypochlorite's

• it has broad spectrum antimicrobial


effect
• inexpensive and fast acting,
• It is recommended for disinfection of
counter tops and work surfaces,
Chlorine
• It is very effective against vegetative bacteria and
viruses like HBV and HIV viruses, therefore its
use is recommended at 1:10 dilution for
disinfection of blood spills
• not much useful against spores
Phenol

• It effective against bacteria


(especially gram +ve bacteria)
and enveloped viruses.
• They are not effective against non
enveloped viruses and spores.
• This compounds is used for decontamination of
the hospital environment, including laboratory
surfaces.
No Disinfectant is substitute for the
following Procedures
• Hand washing (hand hygiene);
• use of personal protective equipment
(PPE )in the lab
• Environmental cleaning
• Appropriate handling of waste and
• Taking care of yourself (e.g. immunization)
Sterilization

1. Heat sterilization
2. Chemical sterilization
3. Radiation sterilization
Heat Sterilization
It is the simplest, most effective and inexpensive
method.
Types of heat sterilization :
a) Steam sterilization (Autoclaving)
b) Dry heat sterilization
Mechanism of Action
• DRY HEAT • MOIST HEAT
• protein denaturation 1. Protein denaturation &
1. Oxidative damage coagulation
2. Toxic effects of 2. Latent heat liberated
increased levels of when steam condenses on
electrolytes cooler surface
3. Hydrolysis & breakdown
of bacterial proteins
Steam Sterilization (MOIST HEAT )

Advantages:
– good penetration
– maintains integrity of liquids (e.g. Lubricants) due
to the 100% humidity within the chamber.
Disadvantages:
– Non stainless steel metal items corrode
– may damage plastic and rubber items
Dry-Heat Sterilization
• Heating at atmospheric pressure and often use a fan to
obtain uniform temperature by circulation.
• Heat at 180º for half hour , 170º for 1 hr., or 160º C for 2
hrs.
Temperature (°C) Time (minutes)

120 480

140 180

150 150

160 120

170 60

180 30
Dry-Heat Sterilization
• Advantages:
• It is effective and safe for metal instruments because
the process does not dull instrument edges or
rust/corrode the instruments.
• Disadvantages:
– Less reliable than autoclaving

– Many materials do not tolerate


dry heat (as media)
Chemical Sterilization

• The chemical compounds used can be:


a) Gas Sterilization
b) Liquid Sterilization

• Generally, chemical sterilization procedures have


the disadvantages of presenting health hazards to
users (e.g. poisonous, flammable, )
Gas Sterilization
• The gas used in this procedure is ethylene oxide.
• Keep objects in constant atmospheric humidity.
• Heat to temperatures between 30º and 60º C,for a
period of 10 hrs.
• Residual ethylene oxide must be ventilated
• It is used for sterilizing endoscopes and ansthetic
apparatus.
Liquid Sterilization
• Can be performed with buffered glutaric aldehyde.
• Procedure:
– Immerse object in liquid for several hours.
– Rinse with sterile water after end of procedure.
– It used in hospitals for sterilization of surgical
instruments, rubber or plastic equipment
which cannot be heat sterilized
Radiation Sterilization

• Provides effective way of sterilization when


using non ionizing radiation in high doses
(Gamma radiation ).

• it is clean process
• dry process
• the object full exposure from all directions
Non ionizing radiations
• UV RAYS (ultra violet)
are used to sterilize internal Surfaces of safety
cabinets, entry ways, Operation theaters etc.

• IR RAYS ( infra red)


are used for rapid mass sterilization of pre packed
items such as syringes, catheters etc.
Sterilizing Filter

• Aqueous liquids sterilized by forced passage through filter of


porosity small enough to retain any microorganism.

1) Membrane filters:
• Manufactured from variety of polymeric material such as
cellulose diacetate, polycarbonate and polyester, as
disc.

• Membrane made in 2 ways


I. Labyrinthine pore membranes: Bacteria & yeast
II. Capillary pore membranes: For viruses

Filters may be supplied with plastic holders pre sterilized for single
use, or mounted in re-usable holders and fitted to filtration
vessels.
Filtration

• HEPA
removes
microbes
>0.3 µm
• Membrane
filtration
removes
microbes
>0.22 µm
Figure 7.4
Sterilizing Filter
2)Syringe filters:

• Membrane 13-25mm diameter.


• Fitted in syringe like holders of stainless steel or
polycarbonate.
• Used for sterilization of small volumes of fluid.

3)Vacuum and in- line filters:

• Membranes of 25-45 mm diameter are used either


with in line filter holders of Teflon or stainless steel
and aluminum.
• Used for sterilization of large volumes of air and
liquid.
Asepsis

Sterilization Disinfection

Heat Chemical Radiation Heat Chemical

Steam Gas Gamma Boiling water Phenol

Dry heat Ethylene oxide 70% Alcohol

Formalin Hexachlorophene

Heat or
Liquid
chloramines solution

Glutaric
Aldehyde
In summary

• Sterilization and disinfection are costly and time


consuming process
• However, it is an essential in all health care
facilities to avoid infection.
• It depends the material and users of instrument.
• Staff should keep in mind that contaminated
instrument present risk to the sample as well as
the user himself.
30

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