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Bacteriology > Direct Detection Methods > Flashcards

Direct Detection Methods Flashcards

1
Q

ENUMERATE THE ROLE OF MICROSCOPY

A

To determine presence of an infection

To characterize organisms grown in culture

To visualize etiologic agents

To screen for specimen adequacy

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2
Q

is used to check if sputum is adequate for culture, whether it is just pure mucus or actually sputum itself

A

Bartlett’s classification

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3
Q

materials known or reasonably expected to contain a pathogen

A

Etiologic agents (infectious substances),

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4
Q

These are examples of?

(i.e., bacteria, yeast, parasites) not readily seen by the naked eye alone

A

Etiologic agents (infectious substances),

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5
Q

A visible light is passed through the specimen – specimen’s image appears dark against a brighter background

A

BRIGHT-FIELD (LIGHT) MICROSCOPY

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6
Q

extent to which the focused object is maintained in microscopy

A

Resolution

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7
Q

the closest distance between two objects that allows to be distinguished from each other when magnified in microscopy

A

Resolving Power

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8
Q

Allows the object to stand out from the background. Commonly achieved thru staining techniques

A

Contrast

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9
Q

is what is used to visualize bacteria. It is also the only objective lens that requires a substance to visualize slides, as the substance (oil) prevents the scattering of light to visualize the outlines of the specimen

A

The Oil Immersion Objective

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10
Q

Has no specific distance as it is dependent on your own vision

A

Interpupillary Distance

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11
Q

The ability of the microscope to not lose focus when switching objectives

A

Parfocal

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12
Q

Direct observation of unstained specimens – wet mount / wet preparation

A

PHASE-CONTRAST MICROSCOPY

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13
Q

Direct examines of organism , NO STAINING

A

Wet Mount

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14
Q

Uses a SPECIAL CONDENSER is used with an annular (ring-shaped) diaphragm – direct and reflected/diffracted light rays are brought together to produce the image

A

PHASE-CONTRAST MICROSCOPY

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15
Q

Light from below the specimen is blocked (condenser), so that it reaches the object at a sharp angle – specimen appears luminous against a dark background

A

DARKFIELD MICROSCOPY

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16
Q

Used primarily to view spirochetes

A

DARKFIELD MICROSCOPY

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17
Q

Microscopy limited to specialized research facility

A

DARKFIELD MICROSCOPY

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18
Q

Absorb light in UV range and emit visible light at a longer wavelength (when returning to their normal/lower energy state

A

Fluorochromes / Fluorophores (fluorescent dyes)

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19
Q

Direct chemical interaction with the component of the bacterial cell (i.e., Acridine orange, Auramine, Calcofluor white)

A

Fluorochroming

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20
Q

antibodies are conjugated (monoclonal antibodies) to a fluorescent dye (i.e., Fluorescein isothiocyanate / FITC

A

Immunofluorescence

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21
Q

Fluorescing object appears bright against a dark background

A

FLUOROSCENT MICROSCOPY

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22
Q

PART OF FLUOROSCENT MICROSCOPY

prevents the excitation wavelengths from damaging the observer’s eyes; selects and limits the wavelength of transmitted light

A

Excitation / Barrier filter

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23
Q

often used to maintain permanent records in FLUOROSCENT MICROSCOPY

A

Digital photography

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24
Q

IN FLUOROSCENT MICROSCOPY

It is the excitation light that activates the Fluorochromes, which result in the production of energy

TRUE OR FALSE

A

TRUE

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25
Q

The process of coloring the microorganism prepared in a thin film on a slide known as smear (sample)

A

STAINING

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26
Q

smear prepared directly from patient sample / specimen

A

DIRECT SMEAR

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27
Q

smear prepared from grown cultures in artificial media (preferably from 24-hour cultures)

A

Indirect smear

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28
Q

Identify the smear being described

 Checks slow-growing organisms
 Used during Antibiotic treatment
 Done when specimen is not processed immediately or
appropriately

A

Indirect smear

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29
Q

2 types of differential staining

A

Gram Staining and Acid Fast Staining

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30
Q

Principal stain utilized in bacteriologic examination

A

Gram Staining

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31
Q

Why is Gram Staining performed

A

To identify the causative agent may it be gram +, gram -, or even yeast.

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32
Q

Provides rapid presumptive identification of pathogens and assessment of specimen quality

A

Gram Staining

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33
Q

In Gram Staining they retain the stain based on the characteristics of their cell wall

A

Bacteria

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34
Q

Cell walls that retain the crystal violet-iodine (CV-I) complex

A

Thick cell walls (peptidoglycan

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35
Q

is commonly negatively charged so all stains used to visualize them are most likely basic dyes at Ph 7

A

Bacteria

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36
Q

Primary stain: stains all bacteria blue to purple

A

Crystal Violet

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37
Q

Mordant: enhances reaction between cell wall and primary stain

A

Gram’s Iodine

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38
Q

Decolorizer: gram-positive bacteria retain the primary stain because of the peptidoglycan and teichoic acid cross-links. Gram-negative bacteria lose the primary stain because of the large amount of lipopolysaccharide in the cell wall

A

Ethyl alcohol or acetone

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39
Q

Counterstain: no effect on gram-positive bacteria; stains gram-negative bacteria pink to red

A

Safranin O or carbolfuchsin

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40
Q

is the most critical step in gram staining

A

DECOLORIZATION

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41
Q

May result in false gram –

A

Overdecolorization

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42
Q

May result in false gram +

A

Underdecolorization

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43
Q

Mordants like gram iodine enhances the binding of crystal violet to thick cell walls of bacteria

TRUE OR FALSE

A

TRUE

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44
Q

Debris like proteins, squamous epithelial cells, polymorphonuclear cells, WBC’s, etc… will stain Gram – (red). Usually Gram + is reserved for yeast and other Gram + organisms

TRUE OR FALSE

A

TRUE

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45
Q

Quantitate the organism in Gram Stain Reporting:

10-20 per oil immersion field

A

Many (4+)

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46
Q

Quantitate the organism in Gram Stain Reporting:

6-10 per oil immersion field

A

Moderate (3+)

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47
Q

Quantitate the organism in Gram Stain Reporting:

3-5 per oil immersion field

A

Few (2+)

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48
Q

Quantitate the organism in Gram Stain Reporting:

Fewer than 10 identified on complete smear

A

Rare (1+)

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49
Q

Quantitate the organism in Gram Stain Reporting:

None were seen

A

None

50
Q

Quantitate the cells (WBC’s, RBC’s and epithelial cells) in Gram Stain Reporting:

25 or more per low-power field

A

Many (4+)

51
Q

Quantitate the cells (WBC’s, RBC’s and epithelial cells) in Gram Stain Reporting:

10-25 per low-power field

A

Moderate (3+)

52
Q

Quantitate the cells (WBC’s, RBC’s and epithelial cells) in Gram Stain Reporting:

2-10 per low-power field

A

Few (2+)

53
Q

Quantitate the cells (WBC’s, RBC’s and epithelial cells) in Gram Stain Reporting:

Fewer than 2 per low-power field

A

Rare (1+)

54
Q

Quantitate the cells (WBC’s, RBC’s and epithelial cells) in Gram Stain Reporting:

None were seen

A

None

55
Q

Designed for bacteria (i.e., mycobacteria) whose cell walls contain long-chain fatty acids (i.e., mycolic acid) – resistant to decolorization with acid-alcohol due to the presence of Mycolic acid

A

ACID FAST STAINING

56
Q

Acid Fast Staining that utilizes heat to allow penetration of primary stain into the cell wall

A

Ziehl-Neelsen (Classic/Hot method)

57
Q

Acid Fast Staining that replaced the heating step with a higher concentration of phenol in the primary stain

A

Kinyoun (Cold method)

58
Q

Acid Fast Staining that replaced the conventional acid alcohol concentration to 1% H2SO4 (intestinal coccidian oocysts). The common concentration for decolorizers is 3%

A

Modified Kinyoun

59
Q

These are only partially acid-fast species

A

Nocardia spp. And Cryptosporidium

60
Q

It just takes a speck of red to tell if a certain bacteria is Acid-fast-positive

TRUE OR FALSE

A

TRUE

61
Q

In Acid fast staining we use distilled water to avoid the contamination of tap water bacillus in the specimen

TRUE OR FALSE

A

TRUE

62
Q

Methylene blue, Carbolfuchsin, Crystal violet, Safranin

These are all simple or differential stains?

A

SIMPLE STAINS

63
Q

Utilized for observation of presence of microorganism and their size, shape, and morphology

A

SIMPLE STAINING

64
Q

Contains metachromatic granules stained by (methylene blue)

A

Corynebacterium diphtheriae

65
Q

Stain used in Negative Staining

A

India Ink / Nigrosin Dye

66
Q

Staining the background with an acidic dye that make cells become colorless as they repel the acidic stain (light colored bodies against a dark background)

What is this type of stain?

A

NEGATIVE STAINING

67
Q

What staining is being described here?

o Demonstration of diffuse capsule
o Can be utilized in the study of bacterial gas vacuoles and
viral morphology

A

NEGATIVE STAINING

68
Q

These are stains used in what staining category?

 Anthony’s, Hiss’s and Gin’s (Crystal violet)
 Nigrosin (Nigrosin dye)
 Welch (Crystal violet)

A

CAPSULE STAIN

69
Q

Under capsule staining what undergoes this process?

Treatment with hot crystal violet solution and followed by a copper sulfate solution rinse

A

Welch (Crystal violet)

70
Q

These are stains used in what staining category?

 Gray (Carbol fuchsin & Tannic acid)
 Leifson (Carbol fuchsin, Tannic Acid & Methylene blue)

A

FLAGELLA STAIN

71
Q

Under flagella staining what undergoes this process?

Unstable colloidal suspension of tannic acid salts are used to form precipitate on the cell walls and flagella, increasing its diameter and allows staining

A

Leifson (Carbol fuchsin, Tannic Acid & Methylene blue)

72
Q

These are stains used in what staining category?

 Fuelgen (Carbol fuchsin)
 Acridine orange

A

NUCLEIC ACID STAIN

73
Q

Under nucleic acid staining what undergoes this process?

o Binds all nucleic acid
o Fluorochrome
o Useful in staining mycoplasma

A

Acridine orange

74
Q

Under nucleic acid staining what is specific for DNA?

A

Fuelgen (Carbol fuchsin)

75
Q

These are stains used in what staining category?

 Dorner (Carbol fuchsin & Nigrosin dye)
 Schaeffer-fulton (Malachite green & Safranin)

A

ENDOSPORE (SPORE) STAIN

76
Q

Under endospore (spore) staining what undergoes this process?

o Observation of intracellular refractile bodies
o Heating and alcohol treatment allows penetration and
decolorization of stain

A

Schaeffer-fulton (Malachite green & Safranin)

77
Q

These are stains used in what staining category?

 Auramine-rhodamine
 Calcofluor white

A

FLUORESCENT STAIN

78
Q

Under fluorescent staining what undergoes this process?

o Binds to the mycolic acid found in mycobacteria

A

Auramine-rhodamine

79
Q

Under fluorescent staining what undergoes this process?

o Used in visualizing yeast
o Binds to the cell wall of fungi
 Or their chitin / cellulose
o May also be used to visualize microsporidia

A

Calcofluor white

80
Q

These are stains used in what staining category?

 Lactophenol cotton blue
 Methenamine silver
 Periodic acid-Schiff

A

FUNGAL STAIN

81
Q

Under fungal staining what undergoes this process?

Made up of lactic acid (preservative), phenol (inhibitory to other fungus), cotton blue (stain–chitin)

A

Lactophenol cotton blue

82
Q

Under fungal staining what undergoes this process?

Stain fungal elements found in tissue samples

A

Methenamine silver

83
Q

Under fungal staining what undergoes this process?

o Preferred stain for direct clinical material
o Stain molecules high in carbohydrate content

A

Periodic acid-Schiff

84
Q

The study of the components and functions of the immune system

o Immune cells, WBC’s, antibodies, compliments

A

IMMUNOLOGY

85
Q

The study of disease diagnosis using antibody measurements in patient’s serum

A

SEROLOGY

86
Q

What is being described?

 proteins/polysaccharides
 recognized by the immune system
 found in cell walls as epitopes

A

Antigen

87
Q

molecule capable of eliciting an immune response

A

Immunogen

88
Q

products of the immune response following interaction with an immunogen

A

Antibody

89
Q

All immunogens are antigens, because all antigens can elicit an immune response

TRUE OR FALSE

A

FALSE

All immunogens are antigens, but not all antigens can elicit an immune response

90
Q

Aggregation of soluble antigens and antibodies

A

Precipitation

91
Q

These are under what concept of precipitation?

 Radial immunodiffusion
 Subjective interpretation and low sensitivity

A

Single Immunodiffusion / Oudin gel diffusion

92
Q

Single Immunodiffusion / Oudin gel diffusion if done in a petri dish

A

Radial immunodiffusion

93
Q

Commonly used in the detection of exoantigens

A

Double Immunodiffusion / Ouchterlony gel diffusion

94
Q

These are examples of?

Systemic fungi (i.e., Blastomyces dermatitidis, Coccidioides immitis, Coccidioides posadasii, Aspergillus spp., Histoplasma capsulatum) and Candida

A

Exoantigens

95
Q

What is being described?

 Most widely-used in the detection of T. pallidum
 Venereal Disease Research Laboratory / VDRL
 Rapid Plasma Reagin / RPR

A

Flocculation

96
Q

Clumping or aggregation of cells or coated particles toward a specific antigen / antibody

A

Agglutination

97
Q

Agglutination wherein Natural carrier molecules (i.e., directly on organism’s surface)

A

Direct Agglutination

98
Q

Identify this type of Agglutination

 Identifies antigens rather than antibodies
 Antigen bound to a variety of particles (i.e., latex beads,
gelatin, erythrocytes)

A

Indirect / Passive agglutination

99
Q

Commonly used in detection of:

 Cryptococcus neoformans in CSF / Serum, Clostridium
difficile toxins A and B
 Presence of beta-hemolytic streptococci, antibodies to
viruses (i.e., rotavirus, CMV, rubella, VZV)

A

Indirect / Passive agglutination

100
Q

Febrile agglutinin test, Widal Test, Weil-Felix Test are all test under what agglutination?

A

Indirect / Passive agglutination

101
Q

What type of agglutination is being described?

 Antibody bound to a carrier particle instead of antigen
 Commonly used in rapid identification of antigens from
Group A & B streptococcus, Staphylococcus aureus,
Cryptococcus neoformans

A

Reverse Passive agglutination

102
Q

Agglutination of Carrier molecules

A

Coagglutination

103
Q

Agglutination that uses a killed or treated Staphylococcus aureus organisms (Cowan I Strain) and Antigen-antibody binding protein (Protein A)

A

Coagglutination

104
Q

Agglutination used commonly used in Identification of Lancefield Groups (A, B, C, D, F, G, N), Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenzae (A-F)

A

Coagglutination

105
Q

Utilizes antibody (monoclonal / polyclonal) conjugated to a fluorochrome (FITC)

A

IMMUNOFLUORESCENCE

106
Q

Antibody conjugated to a fluorochrome added directly with the patient antigen fixed on the slide

A

Direct Fluorescent Antibody

107
Q

Immunofluorescence commonly used in detection of antigens toward Legionella pneumophila and Chlamydia trachomatis

A

Direct Fluorescent Antibody

108
Q

More commonly used than Direct Fluorescent Antibody (DFA)

A

Indirect Fluorescent Antibody

109
Q

Patient antibody is incubated with a known antigen attached to a solid phase and is then washed and added with anti-human immunoglobulin conjugated to a fluorochrome

A

Indirect Fluorescent Antibody

110
Q

IMMUNOFLUORESCENCE Commonly used in Legionella spp., Borrelia burgdorferi, T. gondii, VZV, CMV, EBV capsid antigen & nuclear antigen, HSV 1 & 2, rubella, Mycoplasma pneumoniae, T. pallidum, and several rickettsiae

A

Indirect Fluorescent Antibody

111
Q

is commonly used to identify Treponema pallidum seen in patients screening for syphilis

A

Indirect Fluorescent Antibody

112
Q

May also be called Enzyme Immunoassay (EIA) and Described into two competitive and non-competitive

A

ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA)

113
Q

Antigen / antibody bound to an enzyme – utilization of enzymes as labels

A

ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA)

114
Q

ELISA where the Antibody is attached to a microdilution tray, spherical plastic / metal bead

A

Solid-phase

115
Q

ELISA commonly used in detection of HBsAg & early hepatitis B antigens, HIV p24 protein

A

Solid-phase

116
Q

ELISA that is Flow-through and large surface area (absorbent material pulls the reactants through the membrane) –nitrocellulose, nylon

A

Membrane-bound solid phase

117
Q

ELISA commonly used in detection of antibody towards hepatitis virus, HSV 1 & 2, RSV, CMV, HIV, rubella (IgG & IgM), mycoplasmas, chlamydiae, B. burgdorferi, Entamoeba histolytica

A

Membrane-bound solid phase

118
Q

These are what kind of ELISA?

Slot-blot, Dot-blot assays, Cassette-based membrane-bound ELISA

A

Innovations

119
Q

An assay used for detection of inhibitory effects of antibody to viruses, BUT is replaced by particle agglutination tests

A

NEUTRALIZATION ASSAY

120
Q

Assay commonly used in detection of antibodies toward bacterial toxins, extracellular products (i.e., Streptolysin O, Dnase B)

Generally performed at research laboratories

A

NEUTRALIZATION ASSAY

121
Q

Assay that uses demonstration of antibody presence at the patient sample

A

COMPLEMENT FIXATION ASSAY

122
Q

Assay commonly used in detection of antifungal and antiviral antibodies, BUT is replaced by: IFA and ELISA

A

COMPLEMENT FIXATION ASSAY

Bacteriology (4 decks)

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