L4: DIAGNOSTIC BACTERIOLOGY Flashcards

1
Q

Laboratory Procedures performed in Clinical Microbiology Laboratory

A
  1. Microscopic ID
  2. Culture Isolation and ID
  3. Detection of Ag from the agent using Immunoassays
  4. Molecular Methods
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2
Q

Specimen should be collected from the ____ of infection

A

actual site

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3
Q

Include throat cultures, nasopharyngeal cultures, oral cavity

A

Respiratory Tract

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4
Q

Specimen of choice for LRT

A

SPUTUM

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5
Q

Respiratory Tract.
PMNs: ___
Epith cell: ___

A

PMNs: >25/lpf
Epith cell: <10/lpf

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6
Q

____ are preferred over rectal swab

A

Stool specimen

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7
Q

Culture to diagnose Gastroenteritis

A

Gastrointestinal Tract

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8
Q

should be use to enhance the growth of the
pathogen, while inhibiting Normal Flora

A

Enrichment Media

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9
Q

Requested to diagnose UTI

A

Urine

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10
Q

Urine. Specimen of choice:

A

CLEAN CATCH MIDSTREAM SPECIMEN

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11
Q

Urine. If patient cannot void 🡪 _____

A

CATHETERIZED SPECIMEN

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12
Q

Urine. Infants and Young children 🡪 _____

A

SUPRAPUBIC ASPIRATION

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13
Q

should be performed in all Urine Samples

A

Colony Count

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14
Q

positive urine culture:

A

> 100,000 CFUs/mL

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15
Q

presence of bacteria in the Blood

A

BACTEREMIA

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16
Q

TRANSIENT BACTEREMIA – ____

A

NORMAL FLORA

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17
Q

sporadically discharged from extravascular abscesses into the blood

A

Intermittent Bacteremia

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18
Q

constant release of bacteria into the blood

A

Continuous Bacteremia

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19
Q

Highest conc of bacteria in the blood 🡪 ___

A

Before the fever spikes

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20
Q

2 or 3 venipuncture sites, __ hour apart

A

1 hour apart

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21
Q

Cleaning of the skin prior for collection of blood: ___ Alcohol, ___ Iodine (left 1min)

A

70% Alcohol, 2% Iodine (left 1min)

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22
Q

____ ml blood is collected from adults during Phlebotomy,

A

10 ml

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23
Q

during Phlebotomy, ____ml is collected from from children

A

1-5ml

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24
Q

Cerebrospinal Fluid. Collected by lumbar puncture (____ Lumbar vert)

A

3rd-4th Lumbar vert

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25
Q

Cerebrospinal Fluid. 3 tubes

A

TT1
TT2
TT3

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26
Q

Cerebrospinal Fluid. 3 tubes. TT1🡪___

A

chemistry/immunology

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27
Q

Cerebrospinal Fluid. 3 tubes. TT2🡪___

A

Microbiology

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28
Q

Cerebrospinal Fluid. 3 tubes. TT3🡪___

A

Hematology

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29
Q

Cerebrospinal Fluid Specimen should be screened IMMEDIATELY for the presence of bacteria using ____

A

Gram stain

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30
Q

DELAYS must be avoided in processing CSF result 🡪 _____

A

HIGH MORTALITY

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31
Q

Generally preferred to collect specimens using SYRINGE and NEEDLE to
aspirate (avoid NF and high chance of ANAEROBES)

A

Wound Culture and Abscesses

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32
Q

result from animal bites, burns, ulcer and traumatic wound

A

Exogenous wound

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33
Q

bacterial sources within the patient (cellulitis, dental infections, septic arthritis)

A

Endogenous wound

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34
Q

______ limit: collection 🡪 Lab reception

A

2 hour limit

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35
Q

Specimen Transport. With delay, use _____ (increase the viability of the pathogen up to 72 hours), and refrigerate the specimen

A

STUART’S MEDIUM

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36
Q

For transport of Stool specimen:

A

CARY-BLAIR medium

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37
Q

MORE PREFERRED than cotton swab

A

Ca-Alginate or Dacron swab

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38
Q

Specimen not collected properly and accordingly 🡪 NOTIFY ______

A

PHYSICIAN or NURSE

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39
Q

Upon receipt of the specimen in the lab 🡪 ______ should be done

A

GROSS EXAMINATION

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40
Q

For anaerobic culture 🡪 make sure to use ____

A

ANAEROBIC TRANSPORT MEDIUM

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41
Q

specimen is mixed with saline then view under the microscope

A

Saline Mount

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42
Q

dissolves keratin to make fungal elements more visible

A

KOH preparation

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43
Q

to detect capsules

A

India Ink

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44
Q

Gram Stain primary stain

A

Crystal Violet

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44
Q

capsular swelling, detect capsular Ag

A

Neufeld (quelling) reaction

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45
Q

Gram Stain. Mordant

A

Iodine

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46
Q

Gram Stain. Decolorizing agent

A

Alcohol/Acetone

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47
Q

Gram Stain. Counter stain

A

Safranin

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48
Q

Use to stain Mycobacteria (walls are thick and waxy)

A

Acid Fast Stain

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49
Q

Acid Fast Stain. Hot method:

A

Ziehl Neelsen

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50
Q

Acid Fast Stain. Cold method:

A

Kinyoun

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51
Q

Fluorescent Stains

A

Rhodamine Stains
Acridine orange
Calcofluor white

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52
Q

stain Mycobacteria

A

Rhodamine Stains

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53
Q

useful to demonstrate small amount of
bacteria in blood cultures, CSF, Urthral smears

A

Acridine orange

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54
Q

Fluorescent Stains for fungi

A

Calcofluor white

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55
Q

Mixture of nutrient needed by microorganisms

A

Culture Media

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56
Q

Culture Media. Contain energy providing source:

A

Carbon, Nitrogen, Sulfur, PO4, Oxygen,
Buffers, CHO, Amino acids

57
Q

Media in liquid Form

A

BROTH

58
Q

Media in gel/semisolid; Solidified by using the RED ALGAE ps

A

Agar

59
Q

Types of Culture Media

A
  1. General Isolation Media/ Basic Media
  2. Non selective isolation media/ Enriched Media
  3. Differential media/ Indicator Media
  4. Enrichment Broth
  5. Selective Media
  6. Antibiotic Media
  7. Transport media
60
Q

Aka Supportive media

A

General Isolation Media/Basic Media

61
Q

Support the growth of non fastidious bacteria

A

General Isolation Media/Basic Media

62
Q

General Isolation Media/Basic Media. example

A
  1. Nutrient agar
  2. Trypticase Soy Agar
  3. Nutrient broth
63
Q

Contains a nutrient supplement (blood, serum, egg)

A

Non selective Isolation Media/Enriched Media

64
Q

Use for cultivation of FASTIDIOUS BACTERIA
Use for culturing sterile body fluids

A

Non selective Isolation Media/Enriched Media

65
Q

Enriched Media example

A
  1. Sheep Blood Agar
  2. Chocolate Agar
66
Q

Enriched media that supports the growth of N. gonorrhea, H. influenza

A

Chocolate Agar

67
Q

Provide distinct appearances of microorganisms to aid in their ID. Mostly use to isolate Gr (-) bacteria through the add’n of inhibitory agents against Gr
(+) bacteria.

A

Differential Media

68
Q

Differential Media example

A
  1. MacConkey agar
  2. Eosin-Methylene Blue
69
Q

Differential Media that contains lactose, Bile salts, red indicator and crystal violet

A

MacConkey agar

70
Q

Differential Media that contains lactose, eosin and methylene
blue

A

Eosin-Methylene Blue

71
Q

isolation for Gr (-) Enteric Bacilli

A

MacConkey Agar

72
Q

MacConkey Agar Inhibitor for Gr(+):

A

Crystal Violet, Bile Salts

73
Q

MacConkey Agar. pH Indicator:

A

Neutral Red

74
Q

MacConkey Agar. Color
Acid:
Alkaline:

A

Acid: RED
Alkaline: YELLOW

75
Q

MacConkey Agar. Lactose Fermenter :

A

Pink to Red

76
Q

MacConkey Agar. Non Lactose Fermenter:

A

Colorless

77
Q

Eosin-Methylene Blue Agar. Lactose fermenter:

A

Pink-purple colonies

78
Q

Eosin-Methylene Blue Agar. E. coli:

A

Pink-purple with green metallic sheen

79
Q

Eosin-Methylene Blue Agar. Klebsiella:

A

Pink mucoid colonies

80
Q

Eosin-Methylene Blue Agar. Enterobacter:

A

Pink colonies with dark center (“fish eye colonies”)

81
Q

isolation and differentiation of Vibrio species

A

ThioSulfate Citrate Bile Salts

82
Q

ThioSulfate Citrate Bile Salts (TCBS). pH indicator:

A

Bromthymol Blue

83
Q

ThioSulfate Citrate Bile Salts (TCBS).
ACID:
ALKALINE:

A

ACID: YELLOW
ALKALINE: GREEN

84
Q

ThioSulfate Citrate Bile Salts (TCBS). SF:

A

V. cholera, V. alginolyticus

85
Q

ThioSulfate Citrate Bile Salts (TCBS). NSF:

A

V. parahemolyticus

86
Q

Use to enhance the growth of the bacteria needed. Use frequently for stool specimens to inhibit the normal flora bacteria (E.coli)

A

Enrichment Broth

87
Q

Gram Negative broth (contains ____ )

A

bile salts

88
Q

Selenite Broth (contains Na-H-Selenite) – isolation for ____

A

Shigella, Salmonella

89
Q

Thioglycolate broth (contains ____) – aerobes, anaerobes

A

Thioglycolic acid, nutrients)

90
Q

Solid media that allow one to SELECT for pathogens through the inhibition of normal flora. It selectively favors the growth of a wanted bacteria and inhibit those unwanted

A

Selective Media

91
Q

Contains Salts, CHO, pH indicator, H2S indicator, nutrients

A

Selective Media

92
Q

Selective Media. EX:

A

Hektoen Enteric Agar
Salmonella-Shigella agar
Xylose-lysine-deoxycholate agar

93
Q

Hektoen Enteric Agar. CHO:

A

Lactose, Sucrose, Salicin

94
Q

Hektoen Enteric Agar. H2S indicator:

A

Ferric-NH4-SO4 (black)

95
Q

Hektoen Enteric Agar. LF, H2S (-):

A

Yellow, w/o black center

96
Q

Hektoen Enteric Agar. LF, H2S (+):

A

Yellow, w/ black center

97
Q

-isolation of Salmonella, Shigella

A

Salmonella-Shigella Agar

98
Q

Salmonella-Shigella Agar. pH indicator:

A

Neutral Red

99
Q

Salmonella-Shigella Agar.
ACID:
ALKALINE:

A

ACID: RED
ALKALINE: Yellow

100
Q

Salmonella-Shigella Agar. H2S indicator:

A

Ferric Citrate

101
Q

Salmonella-Shigella Agar.
Salmonella:

A

non LF, colorless with Black Center

102
Q

Salmonella-Shigella Agar. Shigella:

A

Non LF, Colorless w/o black center

103
Q

Agar that is added with antibiotics to selective for certain group of bacteria

A

Antibiotic Agar

104
Q

Antibiotic Agar example

A

Colistin-Nalidixic Acid (CNA)
Thayer Martin Agar
Modified Thayer Martin (MTM)

105
Q

Colistin-Nalidixic Acid (CNA) –
blood agar with;

A

Colistin
Nalidixic acid

106
Q

Thayer Martin Agar - Chocolate agar with

A

Vancomycin
Colistin
Nystatin

107
Q

– inhibits other Gr (+) bacteria

A

Vancomycin

108
Q

– inhibits other gr (-) bacteria

A

Colistin

109
Q

– inhibits growth of yeast

A

Nystatin

110
Q

inhibits swarming of Proteus

A

Trimethopim lactate

111
Q

inhibits fungal growth

A

ANISOMYCIN

112
Q

– prevent fungal growth

A

AMPHOTERICIN B

113
Q

Antibiotic Agar. for N. gonorrhea

A

Martin-Lewis medium

114
Q

Antibiotic Agar. Neisseria

A

New York City Agar

115
Q

Modified Thayer Martin (MTM) –
Chocolate agar with;

A

Vancomycin
Colistin
Nystatin
Trimethoprim lactate

116
Q

Martin-Lewis medium – for N. gonorrhea has:

A

Vancomycin
Colistin
Trimethoprim lactate
ANISOMYCIN

117
Q

New York City Agar – Neisseria. has:

A

Vancomycin
Colistin
Trimethoprim lactate
AMPHOTERICIN B

118
Q

Incubation preferred temperature

A

Most prefer 30-45C

119
Q

Most incubators are set at ____ (preferred by most human internal pathogens)

A

35C

120
Q

Thermophiles:

A

> 40C

121
Q

Psychrophiles:

A

4-20C

122
Q

Culture media should be incubated for at least ?

A

24 hours

123
Q

If no growth after 24 hours 🡪 reincubate for another ____

A

24hours

124
Q

Anaerobic culture: ___ days

A

3-6 days

125
Q

For slow growing 🡪 at least __ hours

A

48 hours

126
Q

bioanalytical methods in which the quantitation of the analyte depends on the reaction of an antigen (analyte) and an antibody

A

Immunoassays

127
Q

rely on the ability of an antibody to recognize and bind a specific
macromolecule in what might be a complex mixture of macromolecules 🡪________

A

IMMUNE COMPLEX

128
Q

Immunoassays example

A

Widal Test, Weil Felix Test
Agglutination test
Enzyme-Linked Immunosorbent Assay (ELISA)
Immunofluorescence

129
Q

solid-phase type of enzyme immunoassay (EIA) to detect the presence of a ligand (commonly a protein) in a liquid sample using antibodies directed against the ligand to be measured

A

Enzyme-Linked Immunosorbent Assay (ELISA)

130
Q

examples include: diagnosis of HIV infection, pregnancy tests, and measurement of cytokines or soluble
receptors in cell supernatant or serum

A

Enzyme-Linked Immunosorbent Assay
(ELISA)

131
Q

Immunofluorescence Dyes:

A

fluorescein isothiocyanate (FITC), rhodamine, and
phycoerythrin

132
Q

Sophisticated laboratory procedures and techniques which utilizes DNA,
RNA or proteins, and a genetic code to IDENTIFY various microorganisms
(virus, bacteria) based from their genetic information

A

Molecular Diagnostics

133
Q

Employs HYBRIDIZATION (interaction between 2 ss n.a. molecules ro
form ds molecule)

A

Molecular Diagnostics

134
Q

Molecular Diagnostics. Some methods used in Microbiology Laboratory

A
  1. BLOTS – Western, Southern, Northern
  2. Polymerase Chain Reaction (PCR)
135
Q

Use to identify proteins and nucleic acid sequences

A

Blots

136
Q

Southern Blotting. Target molecule

A

DNA

137
Q

Western Blotting. Target molecule

A

Protein

138
Q

Northern Blotting. Target Molecule

A

RNA

139
Q

a nucleic acid amplification testing procedure that consists of
denaturing, renaturing, elongating, and amplifying a short segment of DNA or RNA

A

Polymerase Chain Reaction

140
Q

used to make millions to billions of copies of a specific DNA sample
rapidly, amplifying a very small sample of DNA sufficiently to enable
detailed study

A

Polymerase Chain Reaction

141
Q

Extremely sensitive and less labor intensive than Blotting techniques

A

Polymerase Chain Reaction