BIOSAFETY IN MOLECULAR BIOLOGY LABORATORY

Table of content

Molecular biology laboratory Biosafety Principles of Biosafety Risk assessment Standard molecular biological

practices  Laboratory facilities  Safety equipment

MOLECULAR BIOLOGY LABORTARY

Molecular biology laboratory is a place where work is being done to advance understanding of biological processes at the molecular level providing the knowledge needed to solve key problems in human health .

Biosafety The application of knowledge,

techniques and equipment to prevent personal, laboratory and environmental exposure to potentially infectious agents or biohazards.

Principles of Biosafety

A fundamental objective of any biosafety in molecular biology laboratory program is the containment of potentially harmful biological agents.

Principles of Biosafety

Containment The term “containment”

is used in describing safe methods facilities equipment for managing infectious

materials in the laboratory environment where they are being handled or maintained

Principles of Biosafety

Purpose

The purpose of containment is to reduce or eliminate exposure of laboratory workers, other persons, and the outside environment to potentially hazardous agents.

Laboratory Practices and Technique

Persons working with infectious agents or potentially infected materials must be

aware of potential hazards, must be trained and proficient in the practices and

techniques required for handling such material safely

Work and Risk Assessment

The work scope must be defined and the hazards and risks must be assessed before work begins.

risk group classification is to be used for laboratory work only.

Risk group classification

Risk Group 1(no or low individual and community risk)

Risk Group 2 (moderate individual risk, low community risk

Risk Group 3 (high individual risk, low community risk)

Risk Group 4 (high individual and community risk)

Standard Microbiological Practices

 

Bags, lab coats, books etc. should be placed at specified locations

Food Facilities and Eating Eating, drinking, smoking,

handling contact lenses, applying cosmetics, and storing food for human consumption are not permitted in laboratory areas. Food must be stored outside the laboratory area in cabinets or refrigerators designated and used for this purpose.

Prohibited Mouth Pipetting Mouth pipetting is prohibited;

mechanical pipetting devices must be used.

Washing hands

Persons must wash their hands after working with potentially hazardous materials and before leaving the laboratory

Safe handling of sharps Policies for the safe handling of

sharps, such as needles, scalpels, pipettes, and broken glassware must be developed and implemented. Whenever practical, laboratory supervisors should adopt improved engineering and work practice controls that reduce risk of sharps

Precautions Careful management of needles and other sharps are of primary

importance. Needles must not be bent, sheared, broken, recapped, removed from disposable syringes, or otherwise manipulated by hand before disposal.

Used disposable needles and syringes must be carefully placed in conveniently located puncture-resistant containers used for sharps disposal.

Non-disposable sharps must be placed in a hard walled container for transport to a processing area for decontamination, preferably by autoclaving.

Broken glassware must not be handled directly. Instead, it must be removed using a brush and dustpan, tongs, or forceps. Plastic ware should be substituted for glassware whenever possible.

Surface and EquipmentDecontamination

The work area should be cleaned and maintained in a sanitary condition.

• Surfaces or equipment where work with biological materials is conducted should be routinely decontaminated.

Waste Decontamination and Disposal

Dispose of all biological waste into appropriate receptacles (Orange Biohazard bags).

Continued…

Live cultures can be treated with Clorox bleach or autoclaved.

Do not toss out into regular trash or down drains without

autoclaving.

Autoclave Sterilization and Safety

sterilize materials or equipment and ensure operator safety

Electricity

The voltages used for electrophoresis are sufficient to cause electrocution. Cover the buffer reservoirs during electrophoresis. Always turn off the power supply and unplug the leads before removing a gel.

Laboratory Facilities

Sink for hand washing Laboratories must

have a sink for hand washing

Emergency Eyewashes

An eyewash station must be readily available in the laboratory

Biosafety manual

Laboratory-specific biosafety manual must be prepared and adopted as policy. The biosafety manual must be available and accessible.

Labels and Signs

Biological materials, agents, waste, potentially contaminated items, and laboratory rooms must be properly identified with labels, signs, or colors. Identification is needed so that responsibilities, material identities, hazards, or controls are communicated to workers, visitors, and others.

Pest Management

Biosafety level require a program to control pests such as insects and rodents.

Labeled Apparatus

Since you will use common facilities, all solutions and everything stored in an incubator, refrigerator, etc. must be labeled

Material Safety Data Sheets for chemicals

A number of chemicals used in any molecular biology laboratory are hazardous. MSD’s contains information

chemical name, health hazard data, including first aid treatment, physical data fire explosion hazard data reactivity data, spill or leak procedures any special precautions needed

when handling this chemicals.

Doors for access control

Laboratories should have doors for access control

Drains and Disposal

The laboratory should be designed so that it can be easily cleaned. Carpets and rugs in laboratories are not appropriate

Control Access

The laboratory supervisor must enforce the institutional policies that control access to the laboratory.

Animals and plants not associated with the work being performed must not be permitted in the laboratory

Security

Laboratory doors should be self-closing and have locks

the hosting of visitors and the issuance of gate passes, badges, and/or keys to control access to the site, building, and/or room based on each individual’s business needs

Storage of infectious materialPotentially infectious materials must

be placed in a durable, leak proof container during

collection handling, processing storage transport within a facility.

Safety Equipment

Safety Equipment

Personal protection safety cabinets Facility Security Plans

Personal protection

Protective laboratory clothing

Protective laboratory clothing is a garment such as a

lab coat Gown smock uniform.

Face protection Face protection is a safety device such

as a face mask, face shield, or other splatter guard worn over all or part of the face to protect the face from injury or exposure to biological agents.

Hand Protection

Hand protection is a glove or other safety device used on the hand to prevent injury to the hand or direct skin contact with biological materials.

EYE PROTECTION

Exposure to ultraviolet light can cause acute eye irritation. Since the retina cannot detect UV light, you can have serious eye damage and not realize it until 30 min to 24 hours after exposure. Therefore, always wear appropriate eye protection when using UV lamps.

Use UV goggles and common sense when working with the UV light box.

Vaccination

The use of vaccines may provide an increased level of personal protection

Laboratory personnel receive appropriate immunizations or tests for the agents handled or potentially present in the laboratory (e.g., hepatitis B vaccine or TB skin testing)Cultures, tissues, specimens of body fluids, or potentially infectious wastes

Biosafety Cabinets All manipulations of infectious materials within the laboratory must be conducted in the biological safety cabinet.

Class I inward airflow protects worker exhaust to outside (w/wo HEPA

filter)

Class II

Protects worker, product, environmentused with aerosol- transmissiblemicroorganisms, tissue culture

Class III

totally enclosed, ventilated, air-tightsuitable for work with BSL3/4 agents

Facility Design

The design of a facility is important in providing a barrier to protect people working inside and outside the laboratory, and to protect people or animals in the community from infectious agents which may be accidentally released from the laboratory.

Design features

specialized ventilation systems to assure directional airflow, air treatment systems to decontaminate or remove agents from exhaust air

controlled access zones

Design features

airlocks at laboratory entrances separate buildings or modules for

isolation of the laboratory.

Facility Security Plans

Incident, Accident, and Emergency Response

This guide provides both emergency and nonemergency telephone numbers

First-aid boxes first-aid boxes provided at strategic

locations

Spill kits

spill kits should be provided

Fire prevention and fire protection

fire alarm system fire detection system fire doors in good order

Transportation and Transfer of Biological Agents

Transportation

Transportation refers to the packaging and shipping of these materials by air, land, or sea, generally by a commercial conveyance.

Transfer

Transfer refers to the process of exchanging these materials between facilities

Protection

Protection is achieved by: Rigorous packaging appropriate labeling documentation of the

hazardous contents training of workers

Working With Human and Other Primate Cells andTissues

reported as resulting from the handling of human and other primate cells, there is a more significant risk to acquiring infection with HBV or HIV from exposure to human blood and other body fluids

Potential Laboratory Hazards.

chemical agents (e.g. solvents and staining reagents), physical agents (e.g. radioisotopes), biological agents (e.g. viruses and bacteria). Many of these substances are hazardous

(toxic, irritant, infectious, and allergenic) and they also include known or suspected carcinogens.

Diseases caused by Potential Laboratory Hazards certain cancers in lymphoma brain thyroid among women Leukemia skin lung

Guidelines for Work With Toxins or hazards

The laboratory facilities, equipment, and procedures appropriate for work with toxins of biological origin must reflect the intrinsic level of hazard posed by a particular toxin as well as the potential risk s inherent in the operations perform ed. If both toxins and infectious agents are used, both must be considered when containment equipment is selected and policies and procedures are written. If animals are used, animal safety practices must also be considered

CONCLUSION

The objective of containment is to confine biohazards and to reduce the potential exposure of the laboratory worker, persons outside of the laboratory, and the environment to potentially infectious agents.