1. BIOSAFETY IN MOLECULAR BIOLOGY
LABORATORY

2. Table of content
 Molecular biology laboratory
 Biosafety
 Principles of Biosafety
 Risk assessment
 Standard molecular biological
practices
 Laboratory facilities
 Safety equipment

3. 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 .

4. Biosafety
 The application of knowledge, techniques and
equipment to prevent personal, laboratory and
environmental exposure to potentially infectious
agents or biohazards.

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

6. 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

7. 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.

8. 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

9. 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.

10. 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)

11. Standard Microbiological
Practices

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

13. Food Facilities and Eating
 Eating, drinking, smoking, handlin
g 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.

14. Prohibited Mouth Pipetting
 Mouth pipetting is prohibited; mechanical
pipetting devices must be used.

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

16. 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

17. 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.

18. Surface and Equipment
Decontamination

19.  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.

20. Waste Decontamination and
Disposal
 Dispose of all biological waste into
appropriate receptacles (Orange Biohazard
bags).

21. Continued…
 Live cultures can be treated with Clorox bleach
or autoclaved.
 Do not toss out into
regular trash or down drains without
autoclaving.

22. Autoclave Sterilization
and Safety
sterilize materials or equipment and ensure
operator safety

23. 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.

24. Laboratory Facilities

25. Sink for hand washing
 Laboratories must have a
sink for hand washing

26. Emergency Eyewashes
 An eyewash station must be readily available
in the laboratory

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

28. 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.

29. Pest Management
 Biosafety level require a program
to control pests such as insects
and rodents.

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

31. 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.

32. Doors for access control
 Laboratories should have doors for access
control

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

34. 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

35. 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

36. Storage of infectious
material
Potentially infectious materials must be placed
in a durable, leak proof container during
 collection
 handling,
 processing
 storage
 transport within a facility.

37. Safety Equipment

38. Safety Equipment
 Personal protection
 safety cabinets
 Facility Security Plans

39. Personal protection

40. Protective laboratory clothing
Protective laboratory clothing is a
garment such as a
 lab coat
 Gown
 smock
 uniform.

41. 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.

42. 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.

43. 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.

44. 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

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

46. Class I
 inward airflow protects worker
 exhaust to outside (w/wo HEPA filter)

47. Class II
Protects worker, product, environment
used with aerosol- transmissible
microorganisms, tissue culture

48. Class III
totally enclosed, ventilated, air-tight
suitable for work with BSL3/4 agents

49. 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.

50. Design features
 specialized ventilation systems to assure
directional airflow, air treatment systems to
decontaminate or remove agents from
exhaust air
 controlled access zones

51. Design features
 airlocks at laboratory entrances
 separate buildings or modules for isolation of
the laboratory.

52. Facility Security Plans

53. Incident, Accident, and
Emergency Response
 This guide provides
both emergency and
nonemergency
telephone numbers

54. First-aid boxes
 first-aid boxes provided at strategic
locations

55. Spill kits
 spill kits should be
provided

56. Fire prevention and fire
protection
 fire alarm system
 fire detection system
 fire doors in good order

57. Transportation and Transfer of
Biological Agents

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

59. Transfer
 Transfer refers to the process of exchanging
these materials between facilities

60. Protection
Protection is achieved by:
 Rigorous packaging
 appropriate labeling
 documentation of the
hazardous contents
 training of workers

61. Working With Human and Other
Primate Cells and
Tissues
 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

62. 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.

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

64. 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

65. 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.