L4: DIAGNOSTIC BACTERIOLOGY Flashcards
1
Q
Laboratory Procedures performed in Clinical Microbiology Laboratory
A
- Microscopic ID
- Culture Isolation and ID
- Detection of Ag from the agent using Immunoassays
- Molecular Methods
2
Q
Specimen should be collected from the ____ of infection
A
actual site
3
Q
Include throat cultures, nasopharyngeal cultures, oral cavity
A
Respiratory Tract
4
Q
Specimen of choice for LRT
A
SPUTUM
5
Q
Respiratory Tract.
PMNs: ___
Epith cell: ___
A
PMNs: >25/lpf
Epith cell: <10/lpf
6
Q
____ are preferred over rectal swab
A
Stool specimen
7
Q
Culture to diagnose Gastroenteritis
A
Gastrointestinal Tract
8
Q
should be use to enhance the growth of the
pathogen, while inhibiting Normal Flora
A
Enrichment Media
9
Q
Requested to diagnose UTI
A
Urine
10
Q
Urine. Specimen of choice:
A
CLEAN CATCH MIDSTREAM SPECIMEN
11
Q
Urine. If patient cannot void 🡪 _____
A
CATHETERIZED SPECIMEN
12
Q
Urine. Infants and Young children 🡪 _____
A
SUPRAPUBIC ASPIRATION
13
Q
should be performed in all Urine Samples
A
Colony Count
14
Q
positive urine culture:
A
> 100,000 CFUs/mL
15
Q
presence of bacteria in the Blood
A
BACTEREMIA
16
Q
TRANSIENT BACTEREMIA – ____
A
NORMAL FLORA
17
Q
sporadically discharged from extravascular abscesses into the blood
A
Intermittent Bacteremia
18
Q
constant release of bacteria into the blood
A
Continuous Bacteremia
19
Q
Highest conc of bacteria in the blood 🡪 ___
A
Before the fever spikes
20
Q
2 or 3 venipuncture sites, __ hour apart
A
1 hour apart
21
Q
Cleaning of the skin prior for collection of blood: ___ Alcohol, ___ Iodine (left 1min)
A
70% Alcohol, 2% Iodine (left 1min)
22
Q
____ ml blood is collected from adults during Phlebotomy,
A
10 ml
23
Q
during Phlebotomy, ____ml is collected from from children
A
1-5ml
24
Q
Cerebrospinal Fluid. Collected by lumbar puncture (____ Lumbar vert)
A
3rd-4th Lumbar vert
25
Cerebrospinal Fluid. 3 tubes
TT1
TT2
TT3
26
Cerebrospinal Fluid. 3 tubes. TT1🡪___
chemistry/immunology
27
Cerebrospinal Fluid. 3 tubes. TT2🡪___
Microbiology
28
Cerebrospinal Fluid. 3 tubes. TT3🡪___
Hematology
29
Cerebrospinal Fluid Specimen should be screened IMMEDIATELY for the presence of bacteria using ____
Gram stain
30
DELAYS must be avoided in processing CSF result 🡪 _____
HIGH MORTALITY
31
Generally preferred to collect specimens using SYRINGE and NEEDLE to
aspirate (avoid NF and high chance of ANAEROBES)
Wound Culture and Abscesses
32
result from animal bites, burns, ulcer and traumatic wound
Exogenous wound
33
bacterial sources within the patient (cellulitis, dental infections, septic arthritis)
Endogenous wound
34
______ limit: collection 🡪 Lab reception
2 hour limit
35
Specimen Transport. With delay, use _____ (increase the viability of the pathogen up to 72 hours), and refrigerate the specimen
STUART’S MEDIUM
36
For transport of Stool specimen:
CARY-BLAIR medium
37
MORE PREFERRED than cotton swab
Ca-Alginate or Dacron swab
38
Specimen not collected properly and accordingly 🡪 NOTIFY ______
PHYSICIAN or NURSE
39
Upon receipt of the specimen in the lab 🡪 ______ should be done
GROSS EXAMINATION
40
For anaerobic culture 🡪 make sure to use ____
ANAEROBIC TRANSPORT MEDIUM
41
specimen is mixed with saline then view under the microscope
Saline Mount
42
dissolves keratin to make fungal elements more visible
KOH preparation
43
to detect capsules
India Ink
44
Gram Stain primary stain
Crystal Violet
44
capsular swelling, detect capsular Ag
Neufeld (quelling) reaction
45
Gram Stain. Mordant
Iodine
46
Gram Stain. Decolorizing agent
Alcohol/Acetone
47
Gram Stain. Counter stain
Safranin
48
Use to stain Mycobacteria (walls are thick and waxy)
Acid Fast Stain
49
Acid Fast Stain. Hot method:
Ziehl Neelsen
50
Acid Fast Stain. Cold method:
Kinyoun
51
Fluorescent Stains
Rhodamine Stains
Acridine orange
Calcofluor white
52
stain Mycobacteria
Rhodamine Stains
53
useful to demonstrate small amount of
bacteria in blood cultures, CSF, Urthral smears
Acridine orange
54
Fluorescent Stains for fungi
Calcofluor white
55
Mixture of nutrient needed by microorganisms
Culture Media
56
Culture Media. Contain energy providing source:
Carbon, Nitrogen, Sulfur, PO4, Oxygen,
Buffers, CHO, Amino acids
57
Media in liquid Form
BROTH
58
Media in gel/semisolid; Solidified by using the RED ALGAE ps
Agar
59
Types of Culture Media
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
Aka Supportive media
General Isolation Media/Basic Media
61
Support the growth of non fastidious bacteria
General Isolation Media/Basic Media
62
General Isolation Media/Basic Media. example
1. Nutrient agar
2. Trypticase Soy Agar
3. Nutrient broth
63
Contains a nutrient supplement (blood, serum, egg)
Non selective Isolation Media/Enriched Media
64
Use for cultivation of FASTIDIOUS BACTERIA
Use for culturing sterile body fluids
Non selective Isolation Media/Enriched Media
65
Enriched Media example
1. Sheep Blood Agar
2. Chocolate Agar
66
Enriched media that supports the growth of N. gonorrhea, H. influenza
Chocolate Agar
67
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.
Differential Media
68
Differential Media example
1. MacConkey agar
2. Eosin-Methylene Blue
69
Differential Media that contains lactose, Bile salts, red indicator and crystal violet
MacConkey agar
70
Differential Media that contains lactose, eosin and methylene
blue
Eosin-Methylene Blue
71
isolation for Gr (-) Enteric Bacilli
MacConkey Agar
72
MacConkey Agar Inhibitor for Gr(+):
Crystal Violet, Bile Salts
73
MacConkey Agar. pH Indicator:
Neutral Red
74
MacConkey Agar. Color
Acid:
Alkaline:
Acid: RED
Alkaline: YELLOW
75
MacConkey Agar. Lactose Fermenter :
Pink to Red
76
MacConkey Agar. Non Lactose Fermenter:
Colorless
77
Eosin-Methylene Blue Agar. Lactose fermenter:
Pink-purple colonies
78
Eosin-Methylene Blue Agar. E. coli:
Pink-purple with green metallic sheen
79
Eosin-Methylene Blue Agar. Klebsiella:
Pink mucoid colonies
80
Eosin-Methylene Blue Agar. Enterobacter:
Pink colonies with dark center (“fish eye colonies”)
81
isolation and differentiation of Vibrio species
ThioSulfate Citrate Bile Salts
82
ThioSulfate Citrate Bile Salts (TCBS). pH indicator:
Bromthymol Blue
83
ThioSulfate Citrate Bile Salts (TCBS).
ACID:
ALKALINE:
ACID: YELLOW
ALKALINE: GREEN
84
ThioSulfate Citrate Bile Salts (TCBS). SF:
V. cholera, V. alginolyticus
85
ThioSulfate Citrate Bile Salts (TCBS). NSF:
V. parahemolyticus
86
Use to enhance the growth of the bacteria needed. Use frequently for stool specimens to inhibit the normal flora bacteria (E.coli)
Enrichment Broth
87
Gram Negative broth (contains ____ )
bile salts
88
Selenite Broth (contains Na-H-Selenite) – isolation for ____
Shigella, Salmonella
89
Thioglycolate broth (contains ____) – aerobes, anaerobes
Thioglycolic acid, nutrients)
90
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
Selective Media
91
Contains Salts, CHO, pH indicator, H2S indicator, nutrients
Selective Media
92
Selective Media. EX:
Hektoen Enteric Agar
Salmonella-Shigella agar
Xylose-lysine-deoxycholate agar
93
Hektoen Enteric Agar. CHO:
Lactose, Sucrose, Salicin
94
Hektoen Enteric Agar. H2S indicator:
Ferric-NH4-SO4 (black)
95
Hektoen Enteric Agar. LF, H2S (-):
Yellow, w/o black center
96
Hektoen Enteric Agar. LF, H2S (+):
Yellow, w/ black center
97
-isolation of Salmonella, Shigella
Salmonella-Shigella Agar
98
Salmonella-Shigella Agar. pH indicator:
Neutral Red
99
Salmonella-Shigella Agar.
ACID:
ALKALINE:
ACID: RED
ALKALINE: Yellow
100
Salmonella-Shigella Agar. H2S indicator:
Ferric Citrate
101
Salmonella-Shigella Agar.
Salmonella:
non LF, colorless with Black Center
102
Salmonella-Shigella Agar. Shigella:
Non LF, Colorless w/o black center
103
Agar that is added with antibiotics to selective for certain group of bacteria
Antibiotic Agar
104
Antibiotic Agar example
Colistin-Nalidixic Acid (CNA)
Thayer Martin Agar
Modified Thayer Martin (MTM)
105
Colistin-Nalidixic Acid (CNA) –
blood agar with;
Colistin
Nalidixic acid
106
Thayer Martin Agar - Chocolate agar with
Vancomycin
Colistin
Nystatin
107
– inhibits other Gr (+) bacteria
Vancomycin
108
– inhibits other gr (-) bacteria
Colistin
109
– inhibits growth of yeast
Nystatin
110
inhibits swarming of Proteus
Trimethopim lactate
111
inhibits fungal growth
ANISOMYCIN
112
– prevent fungal growth
AMPHOTERICIN B
113
Antibiotic Agar. for N. gonorrhea
Martin-Lewis medium
114
Antibiotic Agar. Neisseria
New York City Agar
115
Modified Thayer Martin (MTM) –
Chocolate agar with;
Vancomycin
Colistin
Nystatin
Trimethoprim lactate
116
Martin-Lewis medium – for N. gonorrhea has:
Vancomycin
Colistin
Trimethoprim lactate
ANISOMYCIN
117
New York City Agar – Neisseria. has:
Vancomycin
Colistin
Trimethoprim lactate
AMPHOTERICIN B
118
Incubation preferred temperature
Most prefer 30-45C
119
Most incubators are set at ____ (preferred by most human internal pathogens)
35C
120
Thermophiles:
>40C
121
Psychrophiles:
4-20C
122
Culture media should be incubated for at least ?
24 hours
123
If no growth after 24 hours 🡪 reincubate for another ____
24hours
124
Anaerobic culture: ___ days
3-6 days
125
For slow growing 🡪 at least __ hours
48 hours
126
bioanalytical methods in which the quantitation of the analyte depends on the reaction of an antigen (analyte) and an antibody
Immunoassays
127
rely on the ability of an antibody to recognize and bind a specific
macromolecule in what might be a complex mixture of macromolecules 🡪________
IMMUNE COMPLEX
128
Immunoassays example
Widal Test, Weil Felix Test
Agglutination test
Enzyme-Linked Immunosorbent Assay (ELISA)
Immunofluorescence
129
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
Enzyme-Linked Immunosorbent Assay (ELISA)
130
examples include: diagnosis of HIV infection, pregnancy tests, and measurement of cytokines or soluble
receptors in cell supernatant or serum
Enzyme-Linked Immunosorbent Assay
(ELISA)
131
Immunofluorescence Dyes:
fluorescein isothiocyanate (FITC), rhodamine, and
phycoerythrin
132
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
Molecular Diagnostics
133
Employs HYBRIDIZATION (interaction between 2 ss n.a. molecules ro
form ds molecule)
Molecular Diagnostics
134
Molecular Diagnostics. Some methods used in Microbiology Laboratory
1. BLOTS – Western, Southern, Northern
2. Polymerase Chain Reaction (PCR)
135
Use to identify proteins and nucleic acid sequences
Blots
136
Southern Blotting. Target molecule
DNA
137
Western Blotting. Target molecule
Protein
138
Northern Blotting. Target Molecule
RNA
139
a nucleic acid amplification testing procedure that consists of
denaturing, renaturing, elongating, and amplifying a short segment of DNA or RNA
Polymerase Chain Reaction
140
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
Polymerase Chain Reaction
141
Extremely sensitive and less labor intensive than Blotting techniques
Polymerase Chain Reaction
