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[MT 6319] Molecular Biology & Diagnostics Laboratory > [1S] UNIT 1.1: Introduction to Molecular Biology and Diagnostics > Flashcards

[1S] UNIT 1.1: Introduction to Molecular Biology and Diagnostics Flashcards

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1
Q

“Field involved in the examination of the origins of diseases at the molecular level, primarily by studying nucleic acids”

A

Molecular Biology & Diagnostics / Diagnostic Molecular Biology

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2
Q

Molecular Biology and Diagnostics is also called as?

A

Diagnostic Molecular Biology

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3
Q

Studies the behavior of living cells in terms of their morphology, anatomy and physiology

A

Cell Biology

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4
Q

Techniques used in Cell Biology

A

Microscopy, Scanning, etc.

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5
Q

Importance: identifying the behavioral patterns of a cell and its morphological features in response to various metabolic, environmental and chemical exposures

A

Cell Biology

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6
Q

Deals with the understanding of the relationship between DNA, RNA and proteins

A

Molecular Biology

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7
Q

Techniques used in Molecular Biology

A

Gel electrophoresis, Polymerase chain reaction, cloning techniques, Sequencing, etc.

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8
Q

Importance: Confirmation of most morphological and metabolic conditions. It is also an important diagnostic tool

A

Molecular Biology

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9
Q

Containment principles technologies and practices implemented to prevent unintentional exposure to pathogens and toxins or their accidental release

A

Biosafety

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10
Q

Protocols and procedures that we do so that the things that we analyze will not be taken out from the laboratory grounds

A

Biosafety

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11
Q

● An agent of biological origin that can cause disease in humans
● It could be the ff:
○ Microorganisms
○ Viruses
○ Allergens or Toxins

A

Biological Hazard/Biohazard

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12
Q

Primary specimen for Molecular Biology

A

Blood

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13
Q

BIOSAFETY

Universal Precautions should be followed, that is, ….

A

All blood samples and other body fluids should be treated as potentially infectious for the transmission of blood borne pathogens like HIV, hepatitis, and others.

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14
Q

Applied to blood and body fluids containing visible blood for all patients.

A

Universal Precautions

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15
Q

Apply the principles of work practice controls and exposure controls to all patients and all patient specimens to prevent exposure of workers to potentially infectious agents and to protect patients.

A

Standard Precautions

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16
Q

Donning & Doffing Sequence

A

Donning:
1) Gown
2) Mask
3) Goggles
4) Gloves

Doffing:
1) Gloves
2) Goggles
3) Gown
4) Mask

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17
Q

BIOSAFETY LEVELS

agents are not known to cause disease

A

Biosafety Level 1

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18
Q

BIOSAFETY LEVELS

Use this BSL when working with:
• Well-characterized agents
• Agents that are of minimal hazard to lab personnel and the environment

A

Biosafety Level 1

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19
Q

BIOSAFETY LEVELS

• E . coliJM109, DH5a
• Saccharomycescerevisiae
• Bacillus subtilis

A

Examples of BSL-1 Agents

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20
Q

BIOSAFETY LEVELS: STANDARD WORK PRACTICES

• Use mechanical pipetting devices
• Wash hands frequently
• Minimize splashes and aerosols
• Decontaminate work surfaces daily
• Handle wastes properly
• Maintain insect and rodent control program

A

BSL-1

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21
Q

PPE for BSL-1

A

• Lab coat or apron
• Safety glasses or goggles
• Gloves as needed

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22
Q

BIOSAFETY LEVELS

agents are associated with human disease

A

BSL-2

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23
Q

BIOSAFETY LEVELS

Use this when working with agents of moderate potential hazard to personnel and the environment

A

BSL-2

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24
Q

BIOSAFETY LEVELS AGENTS

• Human blood or body fluids
• E . coli0157:H7
• Clostridium botulinum
• Retroviral vectors
• Human cells in cell culture

A

BSL-2

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25
Q

BIOSAFETY LEVELS FACILITY

• Adequate illumination
• Eyewash facility
• Negative air pressure
• Autoclave available
• Biological safety cabinet
• Lab must be separated from public areas

A

BSL-2

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26
Q

Are red in color, marked with biohazard symbol, leak proof & puncture resistant

A

Sharps Container (BSL-2 Special Work Practices)

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27
Q

BIOSAFETY LEVELS SPECIAL WORK PRACTICES

• Place used slides and coverslips in sharps containers, never in any other receptacle.
• Sharps containers

A

BSL-2

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28
Q

PPE for BSL-2

A

• Lab coat or apron
• Safety glasses or goggles
• Gloves
• Biosafety cabinet

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29
Q

Aerosols or splashes; Large volumes; High concentrations

A

Biosafety Cabinet

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30
Q

BIOSAFETY LEVELS AGENTS

• HIV
• H1N1 Flu
• Tuberculosis
• Rabies
• Yersiniapestis
• West Nile Virus
• SARS-Cov2

A

BSL-3

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31
Q

BIOSAFETY LEVELS

agents are associated with human disease and are potentially transmitted as aerosols

A

BSL-3

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32
Q

BIOSAFETY LEVELS FACILITY

• Controlled access
• Physical separation from access corridor
• Anteroom; two-self closing interlocked door
• Directional airflow with pressure
• HEPA filtered exhaust
• Change room and shower
• No windows;
• Autoclaves available
• Seamless floors
• Chemical resistant fixtures

A

BSL-3

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33
Q

BIOSAFETY LEVELS SPECIAL WORK PRACTICES

Specific training in handling agents

A

BSL-3 & 4

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34
Q

PPE for BSL-3

A

• Specialized clothing
• Respiratory protection (masks)
• Gloves
• Biosafety cabinet

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35
Q

BIOSAFETY LEVELS

For agents of life threatening nature

A

BSL-4

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36
Q

BIOSAFETY LEVELS AGENTS

• EBOLA
• Small pox

A

BSL-4

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37
Q

BIOSAFETY LEVELS FACILITY

• Controlled access
• 2 laboratory providing absolute separation of the worker from the infectious agents (suit laboratory and cabinet laboratory)
• Filtered air
• Physical separation from access corridor
• Directional airflow with pressure
• Special waste
• Airlock entry

A

BSL-4

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38
Q

PPE for BSL-4

A

• Lab coat or apron
• Positive Pressure suits
• Double ended autoclave
• Biosafety cabinet Class III

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39
Q

T/F: Current WHO guidelines no longer recommend the prescriptive use of Biosafety Levels. Instead, biocontainment are based on 3 categories of requirements anchored on a robust and site-specific risk assessment. However, there are still existing guidelines that still use BSLs as its phaseout is fairly recent

A

T

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40
Q

○ To protect user from the sample
○ To protect the environment from the sample
○ To protect the samples from external elements

A

Biosafety Cabinets

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41
Q

BSCs key aspects

A

• Amount of air exhausted
• Amount of employee protection

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42
Q

BSC CLASS

Personnel & environment

A

Class I

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43
Q

BSC CLASS

Personnel, product, environment; most common in labs

A

Class II A & B

44
Q

BSC CLASS

Specialized conditions

A

Class III

45
Q

BSC CLASS: DIRECTION OF AIRFLOW

Inward (≥75)

A

BSC Class I

46
Q

BSC CLASS APPLICATION

Air intake is through the front face of the cabinet; exhaust exits through a HEPA to the outside or into the room

A

BSC Class I

47
Q

BSC CLASS APPROPRIATE FOR USES OF VOLATILE TOXIC CHEM & RADIONUCLIDES

Acceptable if hard ducted

A

BSC Class I

48
Q

BSC CLASS APPROPRIATE FOR USES OF VOLATILE TOXIC CHEM & RADIONUCLIDES

Minute amounts only if thimble connected to exhaust*

A

BSC Class II Type A1 & A2

49
Q

BSC CLASS APPROPRIATE FOR USES OF VOLATILE TOXIC CHEM & RADIONUCLIDES

Limited amounts*

A

BSC Class II Type B1 & Class III

50
Q

BSC CLASS APPROPRIATE FOR USES OF VOLATILE TOXIC CHEM & RADIONUCLIDES

Acceptable

A

BSC Class II Type B2

51
Q

BSC CLASS APPROPRIATE FOR USES OF VOLATILE TOXIC CHEM & RADIONUCLIDES

Acceptable, must be hard ducted to lab exhaust and operated in Type B mode

A

BSC Class II Type C1

52
Q

BSC PROTECTION

Maximum Product, personnel, and environment

A

BSC Class III

53
Q

BSC PROTECTION

Product only

A

Laminar Flow Clean Bench

54
Q

BCS DIRECTION OF AIRFLOW

Outward (100)

A

Laminar Flow Clean Bench

55
Q

BCS DIRECTION OF AIRFLOW

Inward (75)

A

BSC Class II Type A1

56
Q

BSC DIRECTION OF AIRFLOW

Inward (100)

A

BSC Class II Type A2, B1-2, C1

57
Q

BCS APPLICATION

Air intake is through a HEPA filter; 70% of airflow is recirculated to the cabinet work area through HEPA; 30% balance can be exhausted through HEPA back into the room or to the outside through a canopy unit. Plenum is under positive pressure

A

BSC Class II Type A1

58
Q

BCS APPLICATION

Similar to Class II. A1, but has 100 lfpm face velocity and plenums are under negative pressure to room; exhaust air can be ducted to outside through a canopy unit

A

BSC Class II, Type A2

59
Q

BCS APPLICATION

Air intake is through HEPA filter; 30-40% of airflow is recirculated in the cabinet and the remaining 60%-70% is exhausted through a HEPA filter and a dedicated duct to the outside

A

BSC Class II, Type B1

60
Q

BCS APPLICATION

Air intake is through HEPA filter; 100% of the air is exhausted to the outside via a HEPA-filtered hard duct. No air recirculation in the cabinet.

A

BSC Class II, Type B2

61
Q

BCS APPLICATION

This type of cabinet can be operated in different modes depending on whether the worker protection from hazardous vapors is a consideration. The mode can be set to approximate either a Type A or Type B biosafety cabinet.

A

BSC Class II, Type C1

62
Q

BCS APPLICATION

Totally enclosed, gas-tight, glove ports for manipulation of pathogens. Supply air is HEPA filtered. Exhaust air passes through two HEPA filters in series and is exhausted to the outside via a hard connection. Airflow can be turbulent inside the cabinet

A

BSC Class III

63
Q

BCS PROTECTION

Product only

A

Laminar Flow Clean Bench

64
Q

Laboratory Safety Cabinet Vs Fume Hood and Laminar Flow Center

Closes completely: either horizontally or vertically

A

Chemical Fume Hood

65
Q

Laboratory Safety Cabinet Vs Fume Hood and Laminar Flow Center

  • Not meant for sitting
  • Negative pressure
  • May have solvent/chemical storage underneath
A

Chemical Fume Hood

66
Q

Laboratory Safety Cabinet Vs Fume Hood and Laminar Flow Center

  • Used for strong acids and bases, volatile chemicals
  • Protects the analyst from chemically volatile compounds
A

Chemical Fume Hood

67
Q

Laboratory Safety Cabinet Vs Fume Hood and Laminar Flow Center

  • Fixed sash opening (8 inch.) (alarmed)
  • Sash moves up but does not close completely
  • Designed for seated work
  • Negative pressure - pulls air towards inside the cabinet
A

BSC

68
Q

Laboratory Safety Cabinet Vs Fume Hood and Laminar Flow Center

  • HEPA filter visible in rear or top of unit
  • Usually no sash or sash is fixed
  • Useful for preparation of reagents in PCR
A

Laminar Flow Clean Air Center

69
Q

Laboratory Safety Cabinet Vs Fume Hood and Laminar Flow Center

  • Positive pressure - air blowing into face or breathing zone
  • Protects the samples especially in procedures like amplification or putting samples in the PCR template
A

Laminar Flow Clean Air Center

70
Q

Developed by Occupational Safety and Health Administration (OSHA) for employees who may be exposed to hazardous chemicals in the workplace

A

Hazard Communication Standard – Right to Know Law

71
Q

Need to:
■ Have written hazard communication program
■ Obtain SDSs for each compound
■ Educate employees to interpret labels, SDSs, and health hazards
■ Maintain hazard warning labels

A

Hazard Communication Standard – Right to Know Law

72
Q

● Major source of safety information for employees who may use hazardous materials
● Obtained from manufacturers

A

Safety Data Sheet

73
Q

T/F: In handling biosafety cabinets, you will observe some holes. Avoid blocking those holes. If failed to do so, the BSC will lose its efficiency and will be useless, because it works via the filtration of the air.

A

T

74
Q

substances that affect cellular development of a fetus or embryo

A

Teratogens

75
Q

cancer causing substances

A

Carcinogens

76
Q

chemicals with flashpoint below 37.8°C

A

Flammable

77
Q

chemicals with flashpoint at or above 37.8°C

A

Combustible

78
Q

chemicals injurious to the skin or eyes by direct contact or to the tissue of the respiratory and GIT if inhaled or ingested

A

Corrosive

79
Q

substances that spontaneously explode or ignite or that evolve heat or flammable or explosive gases

A

Reactive

80
Q

Safety when using laboratory equipment, materials, and supplies

A

Mechanical Safety

81
Q

● Direct hazards: death, shock, burns
● Indirect hazards: fire, explosion

A

Electrical Safety

82
Q

ELECTRICAL SAFETY

T/F: Use only explosion-rated (intrinsically wired) equipment in hazardous atmospheres.

A

T

83
Q

ELECTRICAL SAFETY

T/F: Be particularly careful when operating high-voltage equipment, such as electrophoresis apparatus.

A

T

84
Q

ELECTRICAL SAFETY

T/F: Do not use properly grounded equipment (three-prong plug) and do not check for frayed electrical cords.

A

F; use only properly…, check for frayed…

85
Q

ELECTRICAL SAFETY

T/F: Promptly report any malfunctions or equipment
producing a “tingle” for repair. Do not work on “live” electrical equipment.

A

T

86
Q

FIRES

Ordinary combustibles, wood, paper, cloth etc

A

Class A
Pressurized Water & Dry Chemical (PASS)

87
Q

FIRES

Flammable, Liquid, Grease, Gasoline, Paints, Oils,
etc.

A

Class B
Dry Chemical & Carbon Dioxide (PASS)

88
Q

FIRES

Electrical Equipment, Motor Switches

A

Class C
Carbon Dioxide, Halon, Dry Chemical (PASS)

89
Q

FIRES

Flammable Metals, Magnesium

A

Class D
Metal X
Cover burning material with extinguishing agent (scoop, sprinkle)

90
Q

Destroying all microbial life including spores

A

STERILIZATION

91
Q

Destroying majority of microbial life; but not necessarily spores

A

DISINFECTION

92
Q

Chemical disinfectants which can be safely applied to the skin or mucous membrane and are used to prevent infection by inhibiting the growth of bacteria

A

ANTISEPTICS

93
Q

TYPES OF STERILIZATION PROCESS

A
  1. Physical Methods
  2. Chemical Methods
  3. Radiation
94
Q

TYPES OF STERILIZATION PROCESS

● ___ heat: red heat; flaming; incineration; hot air oven
● _____ heat: Pasteurization; boiling; autoclave (steam under pressure)
● Using nanopore filter paper

A

Physical Methods
- Dry
- Heat

95
Q

TYPES OF STERILIZATION PROCESS

bleach, ethanol, hydrogen peroxide, ethylene oxide, paraformaldehyde

A

Chemical methods

96
Q

PHYSICAL METHOD

● steam pressure rises inside and when it reaches the desired set level, the safety valve opens and the excess steam escapes
● From this point, the holding period is calculated. The holding period is 121° C at 15 pound pressure for 15 minutes
● When the holding period is over, the heater is turned off and the autoclave allowed to cool till the pressure inside is equal to the atmospheric pressure

A

Autoclaving

97
Q

PHYSICAL METHOD

● A holding period of 160°C for one hour is used to sterilize glassware, forceps, all-glass syringes, swabs etc.
● Glassware should be perfectly dry before being placed in the oven. Test tubes and flasks should be wrapped in paper.
● The material should be arranged so as to allow free circulation of air in between the objects.

A

Hot Air Oven (Dry Heat)

98
Q

PHYSICAL METHOD

● Membrane filters are widely used to remove unwanted microorganisms from liquids such as sera and solutions during preparation of culture media
● Made of cellulose esters and other polymers with an average pore diameter of 0.22 mm

A

Filtration

99
Q

CHEMICAL METHOD: Using 70% Alcohol or 10% Bleach

T/F: Ideal disinfectant should be:

● Wide spectrum of activity and must be effective against all microorganisms
● Have fast action; high penetrating power; stable
● Not corrode metals
● Not causing irritation
● Cheap
● Easily available and easy to handle

A

T

100
Q

TYPE OF STERILIZATION

● Ultraviolet (UV) for sterilization: very high penetrative power that can be highly lethal to all cells including bacteria
● Used for:
• Cleaning surfaces (inoculation hoods)
• Laboratory benches
• Laboratory apparatus

A

Radiation

101
Q

Three conditions for a successful aseptic technique

A
  1. The work area must be disinfected
  2. The transfer instruments or glasswares must be sterilized
  3. The work must be accomplished quickly and efficiently
102
Q

DISPOSAL OF WASTES

To be lined with clear autoclave bags and should be decontaminated inside the biosafety cabinet and/or autoclaved

A

Biohazard containers

103
Q

DISPOSAL OF WASTES

Contaminated culture tubes and glasswares that are for washing and decontamination

A

Decontamination bins

104
Q

DISPOSAL OF WASTES

Should contain sharps, slides, coverslips, broken glasswares; disposable pipettes and Pasteur pipettes

A

Sharps containers

105
Q

DISPOSAL OF WASTES

Non-contaminated materials

A

Trash cans

[MT 6319] Molecular Biology & Diagnostics Laboratory (8 decks)

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