Yr 2 Block 1 Micro- Diagnostic Techniques

Front 1

what are the three types of microscopy?

Back 1

light and flourescent, confocal laser scanning, and electron

Front 2

what are the two culture techniques?

Back 2

streak plating and urine culture

Front 3

what is the smear preparation for staining techniques?

Back 3

mix with water, smear on slide, allow to air dry, heat so it can stick and then flood with stain, rinse and dry.

Front 4

what are the 5 staining techniques?

Back 4

smear preparation, gram stain, acid-fast stain, capsule stain, and endospor stain

Front 5

Gram stain

Back 5

Primary stain- crystal violent, mordant, 95% ethanol (gram positive and negative) , safranin

Front 6

acid fast stain

Back 6

used particularly for mycobacterium (mycolic acid)  Ziehl Neelsen method (red, hcl/alcohol, Methylene blue).

Front 7

capsule stain

Back 7

presence of extracellular capsules. Bacteria absorbed in skim milk medium. Leaves a halo around the bacterial cells. (congo red, crystal violent)

Front 8

endospore stain (negative)

Back 8

use malachite green . Secondary stain- safranin… vegetative cells take up safranin

Front 9

define general purpose for culture media

Back 9

designed to grow most organisms

Front 10

define selective for culture media

Back 10

favor the growth of distinct groups of microorganisms while inhibiting others

Front 11

define differential for culture media

Back 11

distinguishes microorganisms having a defined metabolic activity)

Front 12

give an example of General purpose, selective, and differential for culture media.

Back 12

GP: blood agar medium
selective: mannitol salt agar
DF: simmon citrate agar

Front 13

PEA

Back 13

Phenylethyl alcohol agar: inhibits gram negative organisms

Front 14

Mannitol-salt

Back 14

 mannitol-salt: indicative of staphylococcus aureus (pH changes the area around the colony yellow). Staph ferments mannitol. Selective for Staphylococci, but differential for staph a.

Front 15

MacConkey

Back 15

selects for gram negative bacteria… those that ferment lactose (e coli) turn indicator to bright pink

Front 16

EMB

Back 16

Eosin Methylene blue agar  inhibits gram positive bacteria and favor gram negative enteric. E. coli= lactose fermenter which results in metallic green colonies. (like tinsel)

Front 17

Thayer-martin

Back 17

blood agar base, selective for Neisseria gonorrhoeae, Neisseria meningitidis

Front 18

XLD

Back 18

xylose lysine desoxycholate isolates salmonella and shigella. H2S production, xylose fermentation and lysine decarboxylation distinguishes salmonella from shigella, black precipitate

Front 19

Hektoen-Enteric

Back 19

particularly salmonella and shigella. contains lactose and determines the fermenters from the non-fermentors. Salmonella produces black precipitates due to thiosulfate. Non-fermentors result in a yellow-orange color in medium.

Front 20

CIN

Back 20

Cefsulodin- irgasan- novobiocin  selective for Yersinia (appears as colonies with red-pigmented bullseye in the middle of each colony). Inhibits gram positive and negative microorganism. 25 degrees C

Front 21

tinsdale

Back 21

isolates corynebacterium diphtheria.--> black precipitates

Front 22

TCBS

Back 22

Thiosulfate citrate-bile salts agar  indicative for vibrio sp

Front 23

BCYE

Back 23

buffered charcoal yeast extract agar  selective for legionella

Front 24

egg yold agar

Back 24

rich in phospholipids  clostridium sp with lecithinase activity. Opaque zones.

Front 25

lowenstein-jensen

Back 25

egg based medium- isolation of mycobacterium sp. Inhibited by malachite green.

Front 26

Bordet-Gengou

Back 26

culture medium that is selective for bordetella pertussis

Front 27

Name the 9 differential medias we studied.

Back 27

mannitol-salt, MacConkey, EMB, XLD, Hektoen-enteric, TCBS, BCYE, egg yolk agar, and Bordet-Gengou

Front 28

catalase

Back 28

Gram Positive, enzyme breaks down hydrogen peroxide into water and oxygen. If positive- staph, negative- strept. 1st step

Front 29

Coagulase

Back 29

after the colony comes back positive (staph), you use coagulase which has the ability to bind fibrinogen causing agglutination of microorganisms. If positive- staph a.

Front 30

Oxidase

Back 30

for Neisseria detects activity of cytochrome oxidase enzyme. Purple end product is indophenol.

Front 31

Indole

Back 31

if positive- example is e. coli. Detects activity of the tryptophanase enzyme on tryptophan…endproduct indole reacts with Kovacs turning yellow to red color

Front 32

DNase

Back 32

catalyzes the depolymerization of DNA. Contains methyl green dye which binds to polymerized DNA. This results in a clear zone.

Front 33

A disc

Back 33

used to differentiate group A streptococci from other groups of beta-hemolytic streptococci.

Front 34

P disc

Back 34

presumptive test for streptococcus pneumoniae

Front 35

Urease

Back 35

key characteristic of proteus sp. Detects activity of enzyme, urease which hydrolyzes urea releasing the endproduct ammonia. pH change  pale orange to pink

Front 36

Citrate utilization

Back 36

test those with the ability to use citrate as sole carbon source. Solution becomes more basic turning the bromothymol blue indicator from green to a bright blue. Positive: enterobacter cloacae. Negative: Escherichia coli.

Front 37

Bile esculin

Back 37

group D streptococci can hydrolyze esculin (enterococcus). Changes color from orange to black via the byproducts of this hydrolysis reacting with iron salts.

Front 38

oxygen requirements

Back 38

growth patterns. Strict aerobe (at top of test tube), strict anaerobe (at bottom), facultative anaerobe (everywhere), and then a negative control.

Front 39

what are enteric tubes?

Back 39

gut tubes arranged on a smaller scale with different testing materials in each box. Take a wire and push it through all of the tubes to inoculate all of them.

Front 40

what are API strips?

Back 40

comprise strips, generally containing 20 miniature biochemical tests, and databases.

Front 41

what is the motility test using soft agar deeps?

Back 41

agar consistency of jello in test tube. Stab the test tube agar with an inoculated wire. If it spreads then the organism is motility, if it stays in the groove then it is not.

Front 42

what are the three types of stains associated with fungus?

Back 42

KOH prep and staining with crystal violent and lactophenol cotton blue or calcofluor white, india ink, giemsa staining

Front 43

KOH prep and staining with crystal violent and lactophenol cotton blue.

Back 43

Dissolves keratin and other cellular material leaving the fungal elements intact. Use when studying fungus under direct microscopic examinations. (which is how most of the studying of fungus is done)

Front 44

KOH prep and staining with calcofluor white

Back 44

fluorescent dye which is added to the KOH prep prior to coverslipping. Binds to polysaccharides present in chitin or cellulose. Fungal elements fluoresce apple-green or blue white

Front 45

india ink (negative staining)

Back 45

 Crytococcus neoformans: tested for from cerebrospinal fluid specimens. It is an encapsulated yeast. Budding yeast surrounded by a large, clear zone against a black background.

Front 46

giemsa staining

Back 46

detects: histoplasma capsulatum in blood or bone marrow. Stains yeast a purple-blue surrounded by clear halo.

Front 47

PCR

Back 47

primers (short and complimentary), 4 deoxynucleotide triphosphates, and taq polymerase enzyme (derived from thermos aquaticus). 2x stranded dna is denatured using heat. 2 primers are annealed.

Front 48

PLFA

Back 48

polylipofattyacid analysis: each microorganism or group has a unique phospholipid fatty acid profile which can be used in identification
 Can also be used to determine live versus dead biomass percentages