ASCP MLT BOC Microbiology

Question Answer

Autoclave,15 pounds of pressure for 15 minutes, 121°C. Kills

What is Steam Under Pressure sterilization?
spores

4 components of Gram stain Crystal violet, Iodine, 95% ethyl alcohol, and Safranin

yellow and beta hemolytic colonies, catalase and coagulase

S. aureus identification
positive, ferments mannitol

White colonies, nonhemolytic, Non-pathogenic. Cat pos, coag

S. epidermidis
neg, mannitol neg sensitive to novobiocin

White to slightly yellow colonies, nonhemolytic, coag neg, may

S. saprophyticus
ferment mannitol, resistant to novobiocin

Gram pos cocci in pairs, tetrads, or clusters, Pigmented

Micrococcus (yellow, orange, tan), nonpathogenic, cat pos, coag neg,
oxidative, sensitive to bacitracin and lysozyme.

pinpoint colonies, grayish white, translucent, beta hemolytic,

Group A strep
PYR pos, Bacitracin susceptible S. pyogenes

larger than Strep A, bulls eye colonies, diffuse beta hemolysis,

Group B strep
Na Hippurate and CAMP pos, S. agalactiae

Group D strep Slightly opaque, nonhemolytic

Gram pos lancet shaped cocci in pairs, colonies are round,

Strep. pneumoniae translucent, dome shaped when young, central depression
with age. Encapsulated, Bile solubility pos, optochin sensitive

alpha hemolytic, normal flora, resistant to optochin and bile

Strep. viridans
solubility neg

nonhemolytic, hydrolyzes esculin, grows in NaCl broth, PYR

Enterococcus
positive

Gram pos rods with square ends, spore forming, anthrax,

Bacillus anthracis
Aerobic, medusa head colonies, nonhemolytic

Non spore forming GPR, Chinese letter or palisade formation,

C. diphtheriae, black colonies with brown halos on Tindale's
Corynebacteria spp
agar, black/grey colonies on cystine tellurite, cat pos,
nonmotile

GPR pleomorphic, non-spore forming, cat neg, H2S pos,

Erysipelothrix nonmotile, Indole neg, resistant to neomycin, Erysipeloid=skin
infection from handling fish or animals
 GPR pleomorphic, vaginal flora, cat neg, chains of rods in
Lactobacillus
THIO, pinpoint alpha-hep colonies to rough grey colonies

GP coccobacilli, pleomorphic, non-spore forming, meningitis,

Listeria monocytogenes BAP and CNA, tiny colonies with narrow zone of beta
hemolysis, esculin pos, cat pos, motile

GPos, fine branching filaments with fragmentation, slow

Nocardia growing, Grows well on Sabouraud dextrose agar, BCYE,
Marin-Lewis, CNA, acid fast, sulfur granules

Gneg diplococci, grows on choc and selective media,

N. gonorhoeae
Superoxol-pos, Sugar utilization, glucose pos

G neg diplococci, choc and selective media, may grow on BAP

N. meningitis
with increases CO2, sugar utilization, glucose and maltose pos

Gneg diplococci grows on selective media, sugar utilization,

N. lactamica
glucose pos, maltose and lactose slow pos, ONPG pos

Gneg diplococci, grows on BAP, choc, and gonococcal selective

Moraxella catarrhaliz
media, Hockey puck colonies, cat and oxidase pos,

antibiotic appropriate for lower RTi caused by Klebsiella

Imipenem
pneumoniae

Blood c/s from patient with Subacute Bacterial Subculture in Pyridoxal-enhanced medium for Strep. mitis
Endocarditis. Cultured on SBA but after incubation, strain that is dependent on Pyridoxal and causes and "culture-
negative (no growth). What should MLS do negative endocarditis"

Aseptate hyphae with sporangiospore Zygomycetes

Jaw surgery anaerobic gram negative cocci Veilonella

Whooping cough specimen of choice Nasopharyngeal swab

Common error in PCR Nucleic acid contamination

Broad base budding cell Blastomyces

Px with respiratory dss > SBA > beta-hemolytic gram

Bacitracin test
positive cocci

Preferred testing for Legionella Urine antigen testing

Cat bite fever Pasteurella multocida

Cat scratch disease Bartonella henselae

Gram positive bacilli > nonmotile > nonhemolytic >

Bacillus anthracis
catalase positive > spore former

Virus > Stool/feces as specimen of choice Rotavirus

Pharyngitis > kidney biopsy Streptococcus pyogenes

Tuberculosis WBC count Monocytosis PS. found in chronic disease; also in endocarditis

Bacteria with no cell wall causes this disease Walking/ Atypical pneumonia: Mycoplasma pneumoniae

Routine method for Rotavirus detection Enzyme immunoassay (EIA)

"Tap water bacillus"; laboratory contaminant Mycobacterium gordonae

Kidney-shaped, gram negative cocci Neisseria gonorrhoea

Gram positve cocci in chains > bile esculin positive > no Group D Streptococcus needs confirmation: Streptococcus
growth in 6.5% NaCl > PYR positive bovis

Detected by latex agglutination test for Staphylococcus

Clumping factor and Protein A
aureus

Diagnosed with chronic hepatitis B infection High titers of IgM anti-HBC

Bile esculin positive > 6.5% NaCl positive > PYR negative
Leuconostoc
> LAP positive

Purpose of potassium permangate in auramine

Quenching agent
rhodamine

Requires olive oil Malassezia furfur

Mucor No rhizoids

Lifetime marker of hepatitis B infection anti-HBC

Parasite causes autoinfection Strongyloides stercoralis

Sterilization process: steam under pressure 121C for 15 mins

The absence of mature trophozoite and schizont is

P. falciparum
observed amongst which Plasmodium spp.

Trichophyton mentagrophytes (+) vs Trichophyton rubrum

Hair perforation test
(neg)

Transudate non-inflammatory

Test to differentiate Pseudomonas aeruginosa and

Growth at 42C
Pseudomonas putida

HIV test first run: reactive 2nd test run: nonreactive Confirm with western blot

Types of media
Aerobes vs anaerobes

Incubation and storage requirements

Specimen collection and transport

QC

Identification of bacteria/fungi/virus/parasite,
Characteristics Methods Staining Antibiotics

Culture and sensitivity testing

Automation

Biochemical tests

Diagnosis and treatment

antler hyphae Microsporum audouinii

What grows on chocolate agar Haemophilus spp. (requires X and V factor)

Double zone of hemolysis and beta lactamase Clostridium perfringens

Clostridium perfringens. Positive reverse CAMP test using

Double zone bacteria. Confirmation test
Streptococcus agalactiae

Gram negative bacilli > beta-hemolytic > oxidase

Aeromonas (hydrophila)
positive

Gram negative diplococci > OPNG negative Neisserria meningitidis

HTLV- confirmatory test Western blot assay

Hepatits B infection marker found in individuals with a

Anti-HBc or HBcAb
past infection

Patient has walking pneumoniae and is prescribed

Bacteria has no cell wall
penicillin. 2 weeks later, still sick. What happened?

Took a swab sample from a wound and incubated on

three different medias (including anaerobic media). No Swab material inhibited the sample.
growth on media. What happened?

Gram positive cocci resistant to furazolidone Micrococcus

Chopped meat agar (iron and glycerol) needed for

Anaerobes
isolation of?

Differentiating biochemical tests for Yersinia pestis and All Yersinia spp. are nonmotile at 37C but motile at 25C
Yersinia enterocolitica Yersinia pestis is nonmotile at both 37C and 25C
Treponemal test with highest specificity and sensitivity Fluorescent treponemal antibody absorption (FTA-ABS) test

Immunity test for Cytomegalovirus (answer needs

Viral culture
confirmation)

Pseudohyphae "When the production of blastoconidia

continues without separation of the conidia from each other,
Blastoconidia are the beginning of
a pseudohypha, consisting of a filament of attached
blastoconidia, is formed."

Organism that gives off a bleach-like odor in culture Eikenella

This spiral-form organism is seen in urine and cultured

Leptospira interrogans
on Fletcher's media

Klebsiella spp. > mucoid, pink colonies on MacConkey

Klebsiella oxytoca and Klebsiella ornithinolytica
agar > A/AG > indole positive

Gram negative bacilli > indole positive > phenyalanine

Proteus vulgaris Proteus mirabilis but indole negative
deaminase positive

You see a curved gram negative bacilli. It was cultured

from the GI tract of a person with ulcers. What test Rapid urease test for Helicobacter pylori
would you do next to confirm its identity?

A person was successfully treated for syphilis 12 years

ago. However, he has just come in again, worried about
VDRL (answer needs confirmation)
having been re-infected. What would you look for in his
blood?

Mycobacterium xenopi "...is a thermophile (survives in hot

Which of the following species of Mycobacterium might
water systems and natural hot water reservoirs) and resistant
be associated with contamination of the hot water
to common disinfectants enabling it to contaminate
system in large institutions such as hospitals?
laboratory samples and medical devices."

3 pairs of hooklets H. diminuta (answer needs confirmation)

growth on the side and around the tip (answer needs

Rhinocladiella description
confirmation)

Beta-hemolytic, spore-forming bacteria Bacillus spp. Gram positve bacilli, appearing like "box cars"

Bacillus anthracis Gram positve bacilli, flat colonies with

Non-hemolytic, spore-forming bacteria
filamentous projections, "Medusa-head"

Enterobacteriaceae that is MR negative Enterobacter Klebsiella

Test to differentiate between Citrobacter spp. and Citrate utilization test Citrobacter spp. positive Escherichia coli
Escherichia coli negative
Test to differentiate Klebsiella spp. and Enterobacter Motility and Ornithine decarboxylase Klebsiella spp. negative
spp. for both Enterobacter spp. positive for both

Lysine decarboxylase, Arginine hydrolase E. cloacae is lysine

decarboxylase negative, arginine hydrolase positive E.
Differentiate different species of Enterobacter
aerogenes is lysine decarboxylase positive, arginine hydrolase
negative

-both selective and differential -contains bile salts and crystal

violet as selective agents that inhibit the growth of gram
MacConkey Agar
positive organisms -can be used to simplify the isolation of
gram negative organisms from a mixed culture

pH indicator -under acid conditions it turns hot pink -when

Neutral red
alkaline it is tan -when neutral it appears a dusty rose color

If bacteria ferments lactose, pink colonies. In addition, if

Lactose positve on MacConkey Agar enough acid is produced they may precipitate the bile salts
resulting in a cloudy pink ring around the colony.

Gram negative bacteria that do not ferment lactose remain

Lactose negative on MacConkey Agar translucent or the color of the medium. Others may turn
alkaline (tan).

-Battery of 4 very useful tests in the identification of Enterics -

especially important as the distinguishing series between 2
IMViC series
closely related bacteria, Escherichia coli and Entrobacter
aerogenes

Some bacteria have the enzyme tryptophanase Tryptophan +

Indole Test
water ---> (tryptophanase) indole + pyruvate + ammonia

-Kovac's indole reagant -positive test is an unmistakeable

Indole reagant bright red layer floating atop the medium -negative tests vary
from yellow to yellow-green to orange

-MRVP medium contains glucose, the substrate of both the

methyl red and Voges-Proskauer tests -some bacteria convert
Methyl Red test
the glucose to many acids and are known as mixed acid
fermenters -Glucose: acid end product

Methyl red -if the media turns red (pink) the test is MR
Methyl Red reagent
positive -if the media turns yellow, the test is MR negative

-determines ability of bacteria to utilize the butylene glycol

fermentation pathway; specifically we will test for an
Vogues-Proskauer test
intermediate compound in this pathway, acetylmethylcarbinol
(AMC) Glucose---> AMC---> butylene glycol (less acid than MR)

Vogues-Proskauer reagent Alpha-napthol and KOH -Agitate vigorously and repeat the
agitation periodically for 30 mins -pink to red color indicates a
 positive VP test -no color change indicates negative VP test

-substrate for Simmons Citrate Agar: citrate -only

carbohydrate source -if bacteria cannot utilize citrate they will
Citrate Utilization test not grow on this medium -bacteria that can utilize citrate will
incorporate it in aerobic pathways and produce alkaline end
products

-pH indicator is bromthymol blue and is already incorporated

Citrate Utilization reagent into the medium -acid pH: yellow -neutral: emerald green -
alkaline: blue

-Blue color on the slant indicates a positive citrate test -if

Citrate Utilization reaction there is no change in the medium (stays green), the test is
negative

Enterics > Lactose fermenters Citrobacter Escherichia Enterobacter Klebsiella

Shigella Yersinia Proteus Salmonella Morganella Providencia

Enterics > Non-lactose fermenters
Serratia Edwardsiella

Enterics > Non-lactose fermenters > H2S negative Shigella Yersinia

Enterics > Non-lactose fermenters > H2S positive Proteus Salmonella

Enterics > Non-lactose fermenters > nonmotie Shigella

Test to differentiate Shigella species Mannitol Shigella dysenteriae is the only one that is negative

Test to differentiate the Mannitol positive Shigella spp.

Phenyalanine deaminase positive Proteus Morganella Providencia

Phenyalanine deaminase negative > H2S positive Yersinia

Test to differentiate between Providencia and

Citrate (answer needs confirmation)
Morganella

Yersinia enterocolitica is positive Yersinia pestis and Yersinia

Ornithine decarboxylase result among Yersinia spp.
pseudotuberculosis are negative (answer needs confirmation)

Spot indole + oxidase - gram negative rods B-hemolytic. If

E. coli ID
negative hemolysis, do PYR. Negative PYR

Oxidase + G- rod with typical smell. Metallic, rough,

Pseudomonas ID
pigmented or extremely mucoid. Indole -

cyclocerine-cefoxitin-fructose agar (CCFA) selective for C. difficile

deoxycholate agar (DEA) Isolation of Enterobacteriaceae. inhibitory for gram positives.

Xylose Lysine Deoxycholate agar (XLD) xylose lysine deoxycholate agar for selection and
differentiation of Enterobacteriaceae. Shigella is red or yellow.
 Salmonella are yellow then red sometimes with black centers.

Spp positive for phenylalanine deaminase Morganella, Providencia, Proteus