Lesson 4 Flashcards
specimen should be collected from the __________ of infection
actual site
in specimen collection, you need to _______ from adjacent tissues/skin
avoid contamination
the quantity of specimen must be _____ in specimen collection
adequate
observe __________ and use only _________ containers/tubes/syringes/swabs
aseptic technique
sterile
in specimen collection, ______ transport of specimen to the laboratory is needed
prompt
samples must be taken/collected prior to _____________
antibiotic administration
in specimen collection, you need to consider the __________ and ________ of the disease
clinical history
physiology
the time from collecting to plating the specimen should be kept in ___________
minimum
the golden rule in specimen collection is?
proper labelling
what are the specific sites for specimen collection?
- respiratory tract
- gastrointestinal tract
- urine
- blood
- cerebrospinal fluid
- wound culture and abscesses
what is the specimen of choice for lower respiratory tract?
sputum
specimen requirements for specimen collection in respiratory tract includes:
throat cultures
nasopharyngeal cultures
oral cavity
the quality of sputum samplkes is determined by the minimum number of ______________ and ______________ per low power field
squamous epithelial cells
polymorphonuclear leukocytes
quality sputum sample:
PMNs _____________
Epithelial cell ____________
> 25/lpf
<10/lpf
thick and sticky substance
sputum
thin and watery substance
saliva
specimens are collected in the gastrointestinal tract for culture to diagnose ______________
gastroenteritis
when collecting specimen in the gastrointestinal tract, _____________ specimen are preferred over _________
stool
rectal swab
what should be used to enhance the growth of the pathogen, while inhibiting normal flora?
enrichment media
this specimen is requested to diagnose UTI
urine
specimen of choice for urines is?
clean catch midstream specimen
if patient cannot void, we usually use _________
catheterized specimen
for infants and young children, we use ________ in collecting urine specimen
suprapubic aspiration
_______should be performed in all urine samples
colony count
a positive urine culture is ___________
> 100,000 CFUs/mL
it is the presence of bacteria in the blood
bacteremia
it is the normal flora in the blood
transient bacteremia
it is sporadically discharged from extravascular abscesses into the blood
intermittent bacteremia
it is the constant release of bacteria into the blood
continuous bacteremia
when is the highest concentration of bacteria in the blood?
before the fever spikes
for venipuncture if 2 to 3 sites, there should be _________ apart in between
1 hour
what are used in cleaning the skin prior for collection of blood, under 1 min.?
70% alcohol
2% iodine
__________ of blood is collected from adults during phlebotomy and ________ from children
10 mL
1-5 mL
it is collected by lumbar puncture
cerebrospinal fluid
what are the specific vertebraes where we collect the CSF?
3rd - 4th lumbar vertebrae
there are 3 tubes in collecting CSF:
TT1
TT2
TT3
What are they for?
chemistry/immunology
microbiology
hematology
specimen should be screened __________ for the presence of bacteria using Gram stain
immediately
delays must be avoided in processing CSF result to avoid ____________
high mortality
it is generally preferred to collect wound culture and abscess specimens using _____ and ______ to aspirate
syringe
needle
syringe and needles are used for wound culture and abscess to avoid _____ and high chance of ________
necrotizing fasciitis
anaerobes
it is the result from animal bites, burns, ulcer, and traumatic wound
exogenous wound
it is the bacterial sources within the patient like cellulitis, dental infections, septic arthritis
endogenous wound
in specimen transport, we should avoid:
exposure to heat
drying
cold temperature
with delay, use ______ to increase the viability of the pathogen up to 72 hours and refrigerate the specimen
Stuart’s medium
it is more preferred than cotton swab
Ca-Alginate or Dacron swab
when specimen is not collected properly or accordingly, you should________
notify physician or nurse
upon the receipt of the specimen in the lab, ________ should be done
gross examination
for anaerobic culture, make sure to use _______________
anaerobic transport medium
specimen is mixed with saline then view under the microscope
saline mount
it dissolves keratin to make fungal elements more visible
KOH preparation
it is used to detect capsules
India ink
is is the capsular swelling, it detects capsular Ag
Neufeld (quelling) reaction
principles of staining technique
fixation
primary stain
mordant
decolorizing agent
counter stain
primary stain
crystal violet
mordant
iodine
decolorizing agent
alcohol/acetone
counter stain
safranin
it is used to stain mycobacteria
acid fast stain
these bacteria have thick and waxy walls
mycobacteria
it is called acid fast because it resist ________ by acid-alcohol
decolorizing
acid fast stain hot method
Ziehl Neelsen
acid fast stain cold method
kinyoun
what are the fluorescent stains?
rhodamine stains
acridine orange
calcofluor white
it stains mycobacteria
rhodamine stains
it is useful to demonstrate small amount of bacteria in blood cultures, CSF, urthral smears
acridine orange
this stain is used for fungi
calcofluor white
it is the mixture of nutrient by microorganisms
culture media
this contains energy proving source like carbon, nitrogen, sulfur, PO4, oxygen, buffers, CHO, amino acids
culture media
culture media may also contain _____________
antibiotics
it is a culture media in liquid form
broth
media in gel/semi-solid form
agar
agar is solidified by using _______________ also known as (agar agar)
red algae
types of culture media
- general isolation media (basic media)
- non selective isolation media (enriched media)
- differential media (indicator media)
- enrichment broth
- selective media
- antibiotic media
- transport media
it is also known as supportive media
general isolation media (basic media)
it support the growth of non fastidious bacteria
general isolation media (basic media)
examples of general isolation media
nutrient agar
trypticase soy agar
nutrient broth
it contains a nutrient supplement like blood, serum, egg
non selective isolation media (enriched media)
it is used for cultivation of FASTIDIOUS BACTERIA
non selective isolation bacteria (enriched media)
is is used for culturing sterile body fluids
non selective isolation media (enriched media)
examples of non selective isolation media
sheep blood agar
chocolate agar
this is an example of non selective isolation media with 5% defibrinated sheep RBC
sheep blood agar
this is an example of non selective isolation media which is heated or enzyme treated blood agar.
chocolate agar
this agar is enriched with dextrose, cysteine, vitamin b, ferric nitrate
chocolate agar
this agar supports the growth of Neisseria gonorrhoeae, Haemophilus influenzae
chocolate agar
this provide distinct appearances of microorganisms to aid in their Identification
differential media (indicator media)
this media is mostly used to isolate Gr(-) bacteria through the addition of inhibitory agents against Gr(+) bacteria
differential media (indicator media)
This media maybe added with indicators that change color when acid is produced for fermentation of carbohydrates (CHO)
differential media (indicator media)
examples of differential media
MacConkey agar
Eosin-Methylene Blue (EMB)
this agar contains lactose, bile salts, red indicator, and crystal violet
MacConkey agar
this agar contains lactose, eosin, and methylene blue
eosin-methylene blue (EMB)
MacConkey Agar is for?
isolation for Gr (-) Enteric Bacilli
MacConkey Agar inhibitor for Gr (+)
crystal violet, bile salts
MacConkey Agar carbohydrates (CHO)
lactose
MacConkey Agar pH indicator
neutral red
MacConkey Agar
acid =
alkaline =
red
yellow
MacConkey Agar lactose fermenter =
pink to red
MacConkey Agar non lactose fermenter =
colorless
Eosin Methylene Blue is for?
isolation of Gr (-) enteric bacilli
Eosin Methylene Blue CHO =
lactose
Eosin Methylene Blue lactose fermenter =
pink-purple colonies
Eosin Methylene Blue non lactose =
colorless
Eosin Methylene Blue Escherichia coli
pink-purple with green metallic sheen
Eosin Methylene Blue
Klebsiella
pink mucoid colonies
Eosin Methylene Blue Enterobacter
pink colonies with dark center (fish eye colonies)
Thiosulfate Citrate Bile Salts (TCBS) is for?
isolation and differentiation of Vibrio species
Thiosulfate Citrate Bile Salts (TCBS)
CHO =
sucrose
Thiosulfate Citrate Bile Salts (TCBS)
pH indicator =
Bromthymol Blue
Thiosulfate Citrate Bile Salts (TCBS)
acid=
alkaline=
yellow
green
Thiosulfate Citrate Bile Salts (TCBS)
spore forming (SF) =
Vibrio cholerae
Vibrio alginolyticus
Thiosulfate Citrate Bile Salts (TCBS)
non-spore forming (NSF) =
Vibrio parahaemolyticus
it is used to enhance the growth of the bacteria needed
enrichment broth
it is used frequently for stool specimens to inhibit the normal flora bacteria (Escherichia coli)
enrichment broth
examples of enrichment broth
Gram Negative broth
Selenite broth
Tetrathionate broth
Thioglycolate broth
Enrichment broth that contains bile salts
Gram Negative broth
Enrichment broth that contains Na-H-Selenite and used as isolation for Shigella
Selenite Broth
contains bile salts and is used for the isolation of Shigella and Salmonella
Tetrathionate broth
Enrichment broth that contains thioglycolic acid and nutrients that isolates aerobes and anaerobes
Thioglycolate broth
Solid media that allow one to select for pathogens through the inhibition of normal flora
selective media
it selectively favors the growth of a wanted bacteria and inhibit those unwanted
selective media
it is used for culturing specimen from a site having a normal microbial normal flora to prevent unwanted contaminant overgrowing a pathogen
selective media
it contains salts, CHO, pH indicator, Hydrogen sulfide (H2S) indicator, nutrients
selective media
examples of selective media
Hektoen Enteric agar
Salmonella-Shigella agar
Xylose-Lysine-deoxycholate agar
it selectively inhibit Gr(+) and Gr(-) bacteria and permit isolation of stool pathogens
Xylose-lysine-deoxycholate agar
it also permits growth of Hydrogen sulfide producing bacteria
Xylose-lysine-deoxycholate agar
Hektoen Enteric agar is for?
isolation of Gr(-) Enteric Bacilli
Hektoen Enteric agar inhibitor for Gr(+) =
bile salts
Hektoen Enteric agar CHO=
lactose, sucrose, salicin
Hektoen Enteric agar pH indicator=
Bromthymol blue
Hektoen Enteric agar
acid=
alkaline=
yellow
green/blue
Hektoen Enteric agar
H2S indicator =
Ferric-NH4-SO4 (black)
Hektoen Enteric agar
Lactose Fermenters, H2S (-) =
yellow w/o black center
Hektoen Enteric agar
Lactose Fermenters, H2S (+) =
yellow w/ black center
Hektoen Enteric agar
non LF, H2S (-) =
Hektoen Enteric agar
non LF, H2S (+) =
Salmonella-Shigella agar is for?
isolation of Salmonella and Shigella
Salmonella-Shigella agar
CHO=
lactose
Salmonella-Shigella agar
pH indicator=
neutral red
Salmonella-Shigella agar
acid=
alkaline=
red yellow
Salmonella-Shigella agar
H2S indicator=
ferric citrate
Salmonella-Shigella agar
Salmonella (non LF)=
colorless with black center
Salmonella-Shigella agar
Shigella (non LF)=
colorless w/o black center
agar that is added with antibiotics to selective for certain group of bacteria
antibiotic agar
examples of antibiotic agar
- Colistin-Nalidixic Acid (CNA)
- Thayer Martin Agar
- Modified Thayer Martin (MTM)
- Martin-Lewis medium
- New York City Agar
an antibiotic agar used for isolation of gram positive bacteria. it is a blood agar with colistin and Nalidixic acid
Colistin-Nalidixic Acid (CNA)
in Colistin-Nalidixic Acid, it disrupts cell membrane, and inhibits other gram negative bacteria
colistin
in colistin-Nalidixic acid, this blocks DNA replication
Nalidixic acid
a type of antibiotic agar that is selective for Neisseria. It is a chocolate agar with Vancomycin, Colistin, and Nystatin
Thayer Martin Agar
in Thayer Martin Agar this inhibits other gram positive bacteria
vancomycin
in Thayer Martin Agar this inhibits growth of yeast
nystatin
it is a type of antibiotic agar which isolates Neisseria. it is a chocolate agar with Vancomycin, Colistin, Nystatin, Trimethoprim lactate
Modified Thayer Martin (MTM)
in Modified Thayer Martin (MTM) this inhibits swarming of Proteus
Trimethoprim lactate
a type of antibiotic agar that is for Neisseria gonorrhoeae. it is composed of vancomycin, colistin, trimethoprim lactate, and anisomycin
Martin-Lewis medium
in Martin-Lewis medium this inhibits fungal growth
anisomycin
it is a type of antibiotic agar that is for Neisseria. It is composed of vancomycin, colistin, trimethoprim lactate, and amphotericin B
New York City Agar
in New York City Agar this prevent fungal growth
Amphotericin B
most preferred mesophilic incubation
30-45 degrees C
most incubators are set @ ________ which is preferred by moist human internal pathogens
35 degrees C
incubation of thermophiles is
.>40 C
incubation of Psychrophiles is
4-20 C
bacteria with oxygen requirement for incubation
obligate aerobes
bacteria that can multiply with/without oxygen
facultative anaerobes
bacteria that can strictly grow in anaerobic environment
obligate anaerobes
bacteria that can grow with 5-10% carbon dioxide with small oxygen
capnophilic
culture media should be incubated for at least _______
24 hours
if no growth after 24 hours, it should be
reincubated for another 24 hours
anaerobic culture takes _______ days
3-6
for slow growing bacteria, it takes at least _______ hours
48
this are bioanalytical methods in which the quantitation of the analyte depends on the reaction of an antigen (analyte) and an antibody
immunoassay
Immunoassay that rely on the ability of an antibody to recognize and bind a specific macromolecules in what might be a complex mixture of macromolecules
Immune complex
examples of immunoassay are:
Widal test
Weil Felix Test
a process in immunology and microbiology where the clumping or aggregation of particles, typically cells or antigens, occurs due to the specific interaction between antibodies and their corresponding antigens.
active agglutination
employs carrier particles that are coated with soluble antigens. In this either antibody or antigen is attached to certain inert carrier thereby, particles or cells gets agglutinated when corresponding antigen or antibody reacts.
passive agglutination
measure the presence of specific antibodies in the sera that inhibit virus-mediated agglutination of erythrocytes.
agglutination inhibition
solid-phase type of enzyme immunoassay to detect the presence of a ligand in a liquid sample using antibodies against the ligand to be measured
Enzyme-Linked Immunosorbent
Assay (ELISA)
it detects and counts certain antibodies, antigens, proteins, and hormones in bodily fluid sample
Enzyme-Linked Immunosorbent Assay (ELISA)
technique used to identify the presence of viral or bacterial antigens and antibodies in a given sample by using a specific dye/fluorescent label which will make the targeted antibodies appear fluorescent
immunofluorescence
immunofluorescence dyes:
fluorescein isothiocyanate (FITC)
rhodamine
phycoerythrin
sophisticated laboratory procedures and techniques which utilizes DNA, RNA, proteins, and a genetic code to identify various microorganisms based on their genetic information
molecular diagnostics
it employs hybridization
molecular diagnostics
some methods of molecular diagnostics that is used in microbiology laboratory
- BLOTS - Western, Southern, Northern
- Polymerase Chain Reaction
it is used to identify proteins and nucleic acid sequences
blots
after isolation the target protein, it transfer to the membrane. what is it?
blots
target molecule of Southern blotting
DNA
target molecule of Western blotting
Protein
target molecule of Northern blotting
RNA
a nuclei acid amplification testing procedure that consists of denaturing, renaturing, elongating. and amplifying a short segment of DNA or RNA
Polymerase Chain Reaction
it is 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
it is extremely sensitive and less labor intensive than blotting techniques
Polymerase chain Reaction
