Brainscape's Knowledge GenomeTM

Browse over 1 million classes created by top students, professors, publishers, and experts.


See full index

1a.)AMT Micro/Para Quick > AMT Micro Quick > Flashcards

AMT Micro Quick Flashcards

1
Q

5 species with capsules:
-mucoid, antigenic/serotype

A

-Cryptococcus neoformans

-Strep/Kleb pneumo
-H.influ, N.meningitis

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

5 nonmotile

Flagella etc
Mono/Amphi/Peri…
Lopho…

A

-Kleb, Shigella

-Bacillus anthracis
-Corneybacterium
-L.mono at 35C

Flagella etc
Mono/Amphi/Peri…flagellus at one,both,all around
Lopho…tuft of flagella at a polar end

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

Suspectibility testing….
-Want most activity, least toxic

-Use Kirby-Bauer method/agar disk diffusion and dilution testing
1.)Make inoculum broth w/4 colonies at 35C 4hrs
-Broth dilution using 0.5 McFarland std adjust turbidity of inoculum
inoculate(acid,barium chloride) 1.5x10^8
2.)Dip sterible swab and streak 3 directions on MH plate
-agar 4.0, MH 150mm, ph 7.2
3.)Wait 5 min, not more 15min to place disk; press down and make sure in contact w/agar(12 disks max)
4.)Incubate 35C, 16-18hrs

Microdilution method:
-Plastic microdilution trays/panels for quantitative MIC
-Wells have micro amts of antimicrobials
-Vitek: agents in wells of plastic cards, optical density is measured
-MicroScan: broth microdilution, manual innoculation, diluted microbes to wells of microdilution tray
-BD Phoenix, TREK

E test is MIC on a plastic strip
-border of growth intersecs strip

A

MBC
minimal bactericidal: 99.9% reduced/killed

MIC
-Read zone with dark back and reflected light
Measure diameter of outer zone with calipers and compare to breakpoints to determine MIC
-mininum inhibitory concentration (lowest concentration ti visibly inhibit growth, lack of visual turbidity)
-MIC is the ability to inhibit multiplication/not kill so it’s bacteriostatic
-MIC done with dilution antimicrobial suspectibility testing

-Compare to breakpoints (diameter of what is suspectable or what is resistant) to measure suspectible, resistant, intermediate(high dose needed)

-smaller zone means too much moisture or too heavy innoculum
-larger zones indicate more antimicrobial activity or too light innoculum
-no zone is complete resistance

Drugs:
Betalactam: ends in ams
-penicillin, cephalosporin, carbapenams
Vancomcin: MRSA, C.diff

QC:
-Plates/Disks stored in fridge, subcultured weekly for disk diffusion
-penicillin/methicillin best indicators of disk storage

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

Resistance: inherent or acquired
- Antimicrobial resistance rates are also strongly correlated with hospital size, tertiary-level care, and facility type
-Enterobacteraceae usually culprits

MDRO:
-Multidrug-resistant organisms (MDROs), including methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE), beta-lactamases (ESBLs)

Beta lactamase:
-made by bacteria, hydrolyzes beta lactam drug, break up beta lactam ring of drug and can’t disrupt bacterias PBP(penicillin binding protein)

-ESBL:
extended sprectrum beta-lactamase; Ecoli, Kleb

-Nitrocefin/Cefinase dist test: yellow to red when beta lactam ring hydrolyzed

D test is double diffusion:
-inducible clindamycin resistance
-looks like a D flattened zone of inhibition to clindamycin
-erythromin can induce clindamycin resistance

A

Resistance cont…

MRSA:
-resistant to oxacillin/methicillin
-nosocomial versus community acquired
-<10mm w/oxacillin disk
-molecular tests for mecA gene
-MRSA is any strain of S. aureus that has developed (through natural selection) or acquired (through horizontal gene transfer) a multiple drug resistance to beta-lactam antibiotics
-altered PBP2a from mec A gene
-MRSA began as a hospital-acquired infection but has become community-acquired, as well as livestock-acquired
-Use vancomycin

VRE
-vanconycin resistant enterococci
-alteration of the peptidoglycan synthesis pathway
-obtain new DNA in the form of plasmids or transposons which encode genes that confer vancomycin resistance
-screen with brainheart infused agar w/vanco

CRE
-carbapenem resistant
-Kleb, Ecoli, Enterobacter
-modified hodge test

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

Describe hemolysis:

alpha prime:
alpha:
beta:
gamma:

A

Describe hemolysis:

alpha prime: aplha surrounded by beta

alpha: incomplete, green

beta: complete, transparent

gamma: no hemolysis

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

QC:
dry heat oven:
autoclave/wet:

A

QC:
dry heat oven: B.subtilis

autoclave/wet: B.stearothermophilus

-dry=shorter bacillus
-wet=longer bacillus

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

7 sterilization methods…
1 most effective…
1 used if media can’t be heated…

A

6 sterilization methods…

-1 most effective…autoclave
-1 used if media can’t be heated…filtration

-boiling
-dry heat/flame
-fractionalization

-glutarldehyde: cold sterilization
-ethylene oxide gas

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

3 antiseptics: tissue

3 disinfectants: surfaces
-one best for viruses,spores,TB,fungi

A

antiseptics:
-alcohol, iodine, hydrogen peroxide

disinfectants:
-chlorine best for viruses, spores
-phenol
-quaternary ammonium

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

Smears:
-thin enough can see individual cells
-2 swabs best: culture, gram
-1 swab only: culture media first then roll swab for gram, slide isnt sterile

A

Morphology: 1
00 objective x 10 ocular=1000x

Cocci
-clusters, chains, pairs, tetrad arrangements
-diplo means pairs
-staphylo means irregular clumps/clusters

Bacilli
-cccobaccilli and fusiform baccilli arrangementes

Spirochetes

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

Stains

-METHYLENE BLUE: simple stain, only one dye

-GRAM: differential stain
-10sec each, rinse in between, air dry, oil objective
1.)crystal violet stains thick peptidoglycan wall with teichoic acid of GP, purple-blue
2.)iodine/mordant: fixes CV in GP; mordant combines with dye forms insoluble complex, fixes CV
3.)alcohol-acetone: decolorizes CV out of GN
4.)safrannin: counterstain for GN, pink-red

-WRIGHT:
*type of Romanowsky stain
*Methanol fixes the cells to the slide.
*Eosin Y is an acidic anionic dye *methylene blue is basic cationic dye
*buffer
*acid components of the cell (nucleus, cytoplasmic RNA, basophilic granules) stain blue or purple
*basic components of the cell (hemoglobin, eosinophilic granules) stain red or orange.

WRIGHT-GIESMA
-Giemsa stain is combined with the Wright’s Stain, the color brightens to a reddish-purple in the cytoplasmic granules

INDIA INK
-Cryptococcus neoformans, because of its large polysaccharide capsule, can be visualized by the India Ink stain.
Organisms that possess a polysaccharide capsule exhibit a halo around the cell against the black background created by the India Ink.
-Nigrosin
-cells negative charge repels stain

A

Acid fast stain

-ACID FAST: mycobacterium/TB
*Hard to decolorize due to mycolic acid and thus retain primary stain
*carbol fuchin, acid alcohol, methylene blue
*Pos pink, Neg blue/green

1.)Kinyoun: cold/no heat
-carbolfuschin
-phenol not heat for penetration
-MB or malachite green counter

2.)ZielNeelson: hot
-carbolfushin primary,
-heat to penetrate
-acid/ethanol to decolorize
-MB counterstain

3.)Fluorochrome:
-dye absorbs UV, emits higher wavelength
-more sensitive
-bright yellow orange, dark back
-auramine/rhodamine primary
-acid alcohol decolorizer
-K permanganate counter

YEAST/FUNGI
-Calcofluor white
-10% KOH to break down debris/mucous
-pos is bluish white flurescence

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

Most abundant NF in throat…

Most common pathogen in throat…

A

NF: alpha heme strep (viridans)

pathogen: S.pyogenes

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

Nasopharygeal swab for these 3…

Throat swab for this…

A

Nasopharygeal: upper respiratory
H.influ, N.mening, B.pertusis

Throat swab:
Strep pyogenes

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

3 GPC in clusters aerobic that are catalase+

4 GPC in chains/pairs aerobic catase -

A

3 GPC in clusters aerobic that are catalase+
-Staph, CNS (epi,sapro), micrococcus

4 GPC in chains/pairs aerobic catase -
-Strep(A pyogens,B agalactiae)
-enterococcus, group D

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

Differentiate CNS S.epi from S.sapro with…
-both cat+, coag neg

S.epi…skin
S.sapro..UTI

A

Differentiate S.epi from S.sapro with…

S.epi…suspectible to Novobiocin

S.sapro..resistant to Novobiocin

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

Grp… strep susceptible to…
How many units…

Grp…strep resistant to…

A

GrpA:
Bacitracin
0.02-0.04

GrpB: resistant to Bacitracin

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

Differentitate Grp A from GrpB strep
-both cat neg

Grp A..
-sensitive to
-1 test pos for

Grp B…
-resistant to
-2 tests are pos for

A

Group A sensitive to Bacitracin
-PYR pos

Group B resistant to Bacitracin
-CAMP, hippurate +

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
17
Q

Both enterococcus and Grp D hydrolyze…

Differentiate these two with…
-enterococcus
-Grp D

A

both hydrolyze esculin

-enterococcus grows in NaCl
-Grp D doesn’t

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
18
Q

What diff GPC aerobic from aerobic GNDC

GPC…
GNDC…

A

oxidase: purple
-drop reagent on colony
-rub colony filter paper, drop reagent
-rub colony on filter with reagent
*but do not use nichrome loop it will give false pos

GPC oxidase neg
GNDC oxidase pos

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
19
Q

Alpha heme strep…

what diff between the two…
S.pneumo…
Viridians

A

S.pneumo:
-bile soluble, optochin susceptible

viridans:
-optochin resistant

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
20
Q

Sugar fermentation…
N.gonn..
N.mening..
N.lac..
Moraxella..
-two test pos for

A

N.gonn=glucose
N.mening=glucose, maltose
N.lac=glucose,maltose, lactose

Moraxella=none
-DNase pos
-Butyrate esterase pos

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
21
Q

2 genera GNDC, oxidase pos…
how to diff..

A

Neiserra, Moraxella

sugars

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
22
Q

Best media for gonococci/meningcocci…
what is main agar, 3 antibiotics, additonal

A

Modified is Thayer Martin plus
-trimethoprim lactate(inhibits proteus swaming)

Thayer Martin
-enriched choc agar
-colistin, nystatin, vancomycin inhibit other bacteria/fungi
-incubate with increase CO2

Martin Lewis: better at inhibiting yeat than TM
NYC
GC-LECT
JEMBEC: transport/growth N.gonn

Chocolate agar:
-enriched, blood lysed supplies X(hemin),V(NAD)
-increased CO2
-Haemopholis/Neisseria

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
23
Q

Two genera that caues pelvic inflammatory disease

A

N.gonn
-no cotton or calcium alginate swab

Chylamydia trachomatis
-no wooden shaft

best in incubate genital for 3 days

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
24
Q

Betalactamase: enzyme by bacteria against beta lactam antibiotics
-do on primary isolation because plasmid coding will be lost by subculturing

-cephalosporin chromogenic color change to pink
-acidometric: turns yellow
-iodometric

A

…

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
25
Q

SALMONELLA AND SHIGELLA
IN STOOL
-Salmonella and Shigella don’t ferment lactose
-Only Salmonella makes H2S
*bile salts to select
*all 3 differentiate LF and H2S
-H2S indicator sodium thiosulfate
*not the same H2S in TSI
(ferric amm.citrate/sulfate)

Hektoen enteric (HE):
-selects for S/S by inhibiting normal GI flora with bile salts, blue/fuschin
-nonpathogens have color
-differentiates LF
-Shigella/Salmonella don’t ferment lactose and don’t change color
-differentiates H2SH2S(NaThiosulfate,Ferric amm.citrate), Salmonella H2S black

Xylose lysine deoxycholate(XLD):
-selects S/S by inhibiting GNR/GP with deoxycholate
-Shigella doesn’t ferment xylose, sucrose, lactose
-differentiates: sugars/LF, H2S(Salmonella black)
-phenol red

Salmonella/Shigella(SS):
-selects by inhibiting other enterics with bile salts, brilliant green
-differentiates: LF, H2S

Gram negative broth:
-S/S from stools/rectal swabs
-selects for S/S by inhibiting GP with deoxycholate and citrate salts

A

…

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
26
Q

In addition to bile salts, each of the 3 have an additional selective agent…

HE:
XLD:
SS:

A

HE: bromothymol blue, acid fuchsin

XLD: deoxycholate

SS: brilliant green

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
27
Q

3 General GN selective media:
-selects for, with, if diff/what diff with/for

EMB:
MAC:
SMAC:

TSI H2S indicators, sugars, reading

A

Eosin Methylene Blue(EMB)
-urine cultures, UTI
-selects for enteric GNR with dyes
-eosin and methylene blue dye inhibit GP
-diff LF green-black to purple (E.coli has green metallic sheen)

MacConkey:
-gastroenteritis
-selects for enteric GN by inhibiting GP with bile salts and crystal violet dye
-diff LF(pink)

Sorbital MacConkey:
-selects for E.coli 0157 with sorbitol, 0157 doesn’t ferment sorbitol and is colorless

Triple sugar agar:
-mostly lactose/sucrose, some glucose
-butt=glucose, slant=sucrose/lactose
-yellow acid, pink=alkalineH2S

TSI indicatiors:
-ferric amm. citrate/sulfate
(not the same is H2S in SS media, Na thiosulfate)
-Salmonella is H2S pos

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
28
Q

Agar: melts at 100, solidifies 45

4 media for GP:

SBA
CNA
PEA
SSA

A

Sheep blood agar:
-selects for nonfastidious, enriched w/blood
-diff with hemolysis/sheep blood

Colistin nalidixic acid:
-selects for GP by inhibiting GN with colistin/nalidixic acid
-sheep blood

Phenylethyl alcohol agar:
-selects for GPC,anaerobic by inhibiting enteric GNR with phenylethyl alcohol

Selective Strep Agar:
-selects for Grp A with antibiotics to suppress NF of throat
-does not different between A or B
-sheep blood

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
29
Q

3 Media for Corynebacterium diptheriae…

which two are diff…what reagent…color…
which one is enrichment…enhances these 2

A

Corynebacterium diptheriae media
*tellurite inhibits NF, C.diph reduce it making it gray-black

Cysteine-tellurite blood agar
-diff w/tellurite, black

Tindale agar:
-diff w/tellurite, gray to black, brown halo

Loeffler
-enrichment for metachromatic granules and pleomorphism

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
30
Q

2 media for Vibrio:

1 select/diff with…color…

A

Thiosulfate citrate bile salts(TCBS)
-diff with sucrose (cholerae is yellow)

Alkaline peptone water(APW)
-enrichment

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
31
Q

2 media for Bordetella pertusis:

Best choice is….

A

Bordetella pertusis

Regan Lowe:
-charcoal agar enriched w/blooed etc

Bordet-Gengou:
-potato glycerol enriched w/blood
-cough plate, mercury droplets colonies

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
32
Q

CIN agar for…colony appears…

BCYE for…has… …

HBT Tween for….

Myco:
Lowenstein-Jensen for…
Middlebrook…

Sabouraud Dextrose Agar, SDA

Slant;
LIA…
TSI…

Urea…

Broths
-Selenite
-Thio

A

CIN: YerSINia, bulls eye colorless halo
*Aero,Plesi, shigelloides

BCYE:
-buffered charcoal yeast extract
-yeast/cysteine enhance Legionella
-charcoal absorbs toxic stuff

HBT Tween: G.vaginalis
-human blood bilayer
-selects for G.vag
-diff w/hemolysis

MYCO TB:
Lowenstein jensen
-myco tb
-egg, potato, glycerol support
-malachite green inhibits
-35C, 5-10% CO2, weekly for 8 weeks

Middlebrook 7H10
-Myco TB

Sabouraud Dextrose Agar, SDA
-fungi
-low ph, 30C

SLANTS:
Lysine Iron Agar, LIA
-lysine, peptones, glucose,
-ferric amm,citrate(H2S, black)
-shows if can decarboxylate or deaminate lysine
-Pos decarboxylate butt is purple
-Pos deaminase slant is burgundy
-Cannot be positive for both

TSI triple sugar iron agar
-must be from pure culture
-isolation of GNR
-sucrose, lactose, sml glucose
-phenol red ph indicator
-H2S thisulfar,ferric amm,citrate

UREA
-utilize urea to ammonia with urease
-phenol red indicator, ph increase magenta

BROTHS:
-Selenite: enrichment for S/S, selenite inhibits
-Thio: anaerobic, decrease O2 tension

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
33
Q

Campylobacter media

broth
-enrichment, subculture to campy selective agar after overnight incubation 4C

Campy BAP
-from stool
-incubate increased CO2 at 42C

A

…

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
34
Q

Anaerobic media

-reducing agents to bind O2 like thioglycollate, cysteine
-broth tubes heated before to drive out O2
-gas paks have Na2CO3 and Na borohydride
-adding water makes H2 and CO2 for growth
-Palladium pellets catalyze the reaction

BBE
-B.fragilis, black

KVLB

CCFA
-C.difficile

Anaerobic PEA
-prevents swarming and enterobac

Chopped/cooked meat

CDC anaerobic blood agar
-hemin, cystine, k enriched

A

…

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
35
Q

5 Gram positive rods aerobic, acronym..

1 random gram positive that is cocoid then 24hrs a rod…

A

BCLEN
-Bacillus
-Corynebacterium
-Listeria monocytogenes
-Erysipelothrix (test tube brush,occupational)
-Nocardia

Rhodococcus equi: coccus to rod 24hrs

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
36
Q

BCLEN: GPR aerobic..

Bacillus…shape, motility

A

Anthracis
spore forming, bamboo pole
string of pearls, Medusa head
nonmotile

cerus:
-food poisoning

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
37
Q

BCLEN: GPR aerobic:

Corynebacterium is a gram…,….

diff from L.mono with… and ….
test for toxin…

NF diptheroid…

Hospital acquired diptheroid

A

Gram positve rod, nonmotile
-chinese letters,picket fence
-pallisade, club shaped
-BabeErnst, metachromatic granules

diff with L.mono:
C.diph is nonmotile/salicin neg
C.diph has toxin, detected with Elek test

NF diphtheroid: pseudodipheriticum

Hospital acquired: jeikeium

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
38
Q

BCLEN: GPR aerobic

Gram postive to gram variable coccobaccilus

Catalase pos

hippurate/camp pos

A

Listeria mono

tumbling motility 25C, cold for months
food poisoning, meningitis newborns

catalase/salicin/motility pos

*looks like GrpB Strep
-but is catalase pos

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
39
Q

BCLEN: GPR aerobic

Both Corynebacterium and Listeria can be gram postive rods aerobic, how to differentiate

A

Listeria is motile and salicin pos

Corynebacterium is nonmotile and salicin neg

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
40
Q

BCLEN: GPR aerobic

GPR aerobic
Fine branching/filaments, beaded
sulfur, urease pos
cannot grow with antibiotics

A

Nocardia

skin=brasil
lung=asteroides

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
41
Q

GNR cause of whooping cough…
prefered media…
alternate medium…

species that is urease/oxidase pos…

A

Bordetella preferss Regan Lowe,
-will grow on Bordet-gengou

bronchiseptica is urease/oxidase pos

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
42
Q

4 GNR fastidious ass with animals

-cat scratch, trench
-cat/dog, musty, bipolar
-goat/cow, undulant fever
-rabbit, tularemia

A

4 GNR fastidious ass with animals

-cat scratch, trench: Bartonella

-cat/dog, musty, bipolar: Pasteurella mul

-goat/cow, undulant fever: Brucella

-rabbit, tularemia: Franciella

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
43
Q

GNR fastidious ass with goat/cow, undulant fever…

-needs CO2, inhibited by thionin
-not on fuchin
-doens’t make H2S

A

GNR fastidious ass with goat/cow, undulant fever…Brucella

-needs CO2, inhibited by thionin: arbortus

-not on fuchin: suis

-doens’t make H2S: melitensis

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
44
Q

GNR fastidious…

-Pontiact fever, AC, BCYE:

-Best on Regan Lowe…two kinds….
-Whooping cough:
-urease/oxidase pos

A

-Pontiac fever, AC, BCYE: Legionella

-Best on regan lowe: Bordetella
-whooping cough: pertusis
-urease/oxidase pos:bronchisepta

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
45
Q

Legionella best medium, second best/color

A

BCYE
-needs cysteine

Feeley Gorman iron cysteine, brown

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
46
Q

2 GNR fastidious, slender, curved
microaerophillic…
how much of each gas

A

Campy: gull winged
Heli

85% N, 10% CO2, 5% O

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
47
Q

3 GNR fastidious, faculative, GI

-rice water stools, TCBS,
string test pos reagent?

-aquatic, doesn’t grow TCBS
-now enterobacteria, doesn’t grow on TCBS

A

3 GNR fastidious, faculative, GI

Vibrio chloreae
-rice water stools, TCBS,
string test pos(0.5% Na deoxycholate)

Aeromonas
-aquatic, doesn’t grow TCBS

Pleisiomonas
-now enterobacteria, doesn’t grow on TCBS

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
48
Q

GNR fastidious, slender, curved
microaerophillic…

-seagulls, 42C, darting corkskrew motility, most common cause of bacterial diarrhea

-doesn’t like 42C, stomach/peptic ulcer, urea breath test

A

GNR fastidious, slender, curved
microaerophillic…

Campy jejuni
-seagulls, 42C, darting corkskrew motility, most common cause of bacterial diarrhea:

Heli pylori
-doesn’t like 42C, stomach/peptic ulcer, urea breath test
-Skirrow agar

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
49
Q

GNCB, pleomorphic
enriched chocolate, fastidious, filamentous
needs X and/or V, satellitism

-upper resp/nasopharyn swab, capsule, needs X,V
-pink eye, needs V
-NF needs mostly V
-Not NF, STD/chancroid, school of fish, needs X

A

Haemophilus:
GNCB, pleomorphic
chocolate, fastidious, filamentous
needs X and/or V, satellitism
-staph supplies V(NAD), hemoylysis releases X(hemin from agar)

influenze:
-upper resp/nasopharyn swab, capsule, needs X,V

aegypticus:
-pink eye, needs V

parainflu,hae,parahae:
-NF needs mostly V

ducreyi:
-Not NF, STD/chancroid, school of fish, needs X

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
50
Q

HACEK: GNCB, Capnophillic, oral flora, endocarditis

A

Haemophilus, Aggre/actino, Cardio,
Eikinella, Kingella

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
51
Q

3 main GNR nonfermenters
-oxidase pos, swimmers ear, grape
-NF skin
-cystic fibrosis/immunocomp, ammonia

A

3 main GNR nonfermenters

Pseudomonas aeruginosa:
-oxidase pos, swimmers ear, grape

Acine-tobacter
-NF skin

Stenotrophomonas maltophilia:
-cystic fibrosis/immunocomp, ammonia

other nonfermenting…
Burk, Elizabeth,Morax, Alcali

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
52
Q

Nonfermentation media, oxidizers….

can see acid production due to…
indicator is…

how may tubes…etc

A

OF oxidation fermentation
-need oxygen, low peptone media
-bromthymol blue indicator
-2 tubes: one open, one covered w/oil

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
53
Q

5 GN intracellular, bacterial like wall, arthropod vectors

-grows in egg yolk sacs, ticks, typhus pox
-2 with morulae in WBC, ticks
-scrub/bush typhus, mites

A

Rikettsae
5 GN intracellular, bacterial like wall, arthropod vectors

Rikettsia:
-grows in egg yolk sacs, ticks, typhus pox

Ehrlicha/Anaplasma:
-2 with morulae in WBC, ticks

Orienta:
-scrub/bush typhus, mites

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
54
Q

Biochemical Reactions…

Catalase:
-break down on H2O2 to O2/H2O
-diff Staph+, Strep -
-pos is bubbles

Coagulase:
-Rapid slide: bound/clumping factor reacts with fibrinogen
-Tube: free/unbound reacts with rabbit plasma forming fibrin clot
-4hrs, 35C, extend to 24hrs if needed
-S.aureus from others

PYR
-is a substrate, N,N, reagent
-id enterococcus and grp A beta strep
-5 min, red

Bile solubility
-bile salt causes autolysis, colony disappears/flat
-Id S.pneumo
-Neg clear, Pos turbid w/in 30min

Spot oxidase
-cytochrome oxidase oxidize Kovacs reagent to purple in 10secs
-id Neisseria and Pseudo aeruginosa(pos)

A

Hippurate hydrolysis
-pos is purple

Motility
-Neg is growth along stab
-Pos is blurred stab line

Spot indole
-enzyme break down tryptophan to indole
-filter paper, aldehyde indicator, blue to green
-id E.coli
-diff Proteus

Esculin
-id Enterococci
-bile inhibits GP
-hydrolyze esculin=pos is black

Salt tolerance
-NaCl, grow in presence of
-pos is cloudy=enterococci

Phenylalaine, tryptophan deaminase
-Pos is green

Lysine iron agar: lia
-used with TSI
-enteric pathogens
-35C, 18-24hrs
-acid=yellow, alkaline=purple
-slant: lysine deaminase
-butt: lysine decarboxylase
-also H2S

Triple sugar iron/TSI/Kligler
-GNB, enterobaceriaceae, enteric
-lactose, sucrose, glucose, H2S
-35C 18-24hrs

Urea
-if have urease, hyrolyze urea to ammonia
-Pos is pink/magenta
-diff Proteus species
-id C.neoformans between other yeasts

Citrate
-use NaCitrate as sole soure of Carbon and amm,phoshate as nitrogen
-enterobacteriaceae
-bromthymol blue: green to blue

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
55
Q

Enterobacteriacea are nonfastidous and…
gram…
oxidase…
ferment…
reduce…

A

gram negative
oxidase neg

ferment glucose
reduce nitrate

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
56
Q

IMViC is…

1st 4 pos is color..
last one pos color…

A

Indole, Methyl red, VP all are red/pink pos

Citrate is blue

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
57
Q

IMViC acrynon and genus:

++–

–++

A

PEE ++–
-Proteus, E.Coli, Edwardsella

KES –++
-Klebsiella, Enterobacter, Serratia

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
58
Q

3 aminos in Decarboxylase test/acrynm…
How many tubes…

A

Decarboxy: LAO (lysine, arginine, orthinine)
4 tubes (one for each, ctrl)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
59
Q

PD(phenylaline deaminase) and urease acronym

A

PD(deaminase)+ urease=PPM
-Proteus
-Providencia
-Morganella

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
60
Q

Urea pos acronym

A

Urea PUNCH
proteus
ureaplasma
nocardia
Cryto neo
Heli

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
61
Q

Nitrate reduction:
-initial pos looks like…
-add zinc pos looks like…

A

initial red, zinc clear is pos

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
62
Q

ONPG tests…
detects…
which organism is ONPG pos

A

slow LF
detects beta galactosidase

citrobacter

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
63
Q

DNAse pos is…

two DNAse pos

A

clearing around colony 0.1N HCl

m.catt, serretia

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
64
Q

Antigen serotyping:
O=
H=
K=

A

O=sOmatic, both salmonella, shigella

H=flah-gella, only salmonella is motile

K=Kapsule, S.typhi

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
65
Q

Acceptable anaerobic..

Unacceptable…

A

-bile, blood, BM, BF
-aspirates
-tissue,wound

unacceptable:
-swabs
-sputum, washings,gastric
-voided, catheterized urine
-feces

66
Q

Anaerobic gas conditions…

random fact: most clinically sig aerobes are faculative anaerobes that can grow in either

A

80-90 N
5-10 CO2
5 H

No oxygen!!!

67
Q

Anaerobic cocci:
1 GPC
1 GNC

A

Anaerobic

GPC: peptococcus
GNC: Veilonella (red fluoresence)

68
Q

Acronym for anaerobic GPR

A

Anaerobic GPR
CLAP

-Clostridium(spores)
-Lacto(vagina pH)
-Actinomyces israelli(lumpy jaw)
-Propionibacterium(acne,NF)

69
Q

2 Clostridium that have oval subterminal spores…

1 has tennis racket terminal spores..
1 has round terminal spores…

A

2 Clostridium that have oval subterminal spores…
-botulinum: canned
-difficile: antimicrobial diarr

1 has tennis racket terminal spores..
-tetani: lockjaw, soil

1 has round terminal spores…
-ramosum

*Perfeingens: double zone hemolysis, box car, gas gangrene

70
Q

4 GNR anaerobic

A

-Bacteriodes fragilis (resistant to penicillan)
-Fuso: tapered, chartreuse
-Prevo, Porphy: brick red

71
Q

Genera that is acid fast, high lipid content,
need 5-10% CO2 at 35C

id off growth, morph, temp, biochem, light rxn, pigment

A

Myco-bacteria

72
Q

Mycobaceteria temp, gas req

NALC:
NaOH:

A

35C
5-10CO2

NALC: liquify
NaOH: digest/decontaminate

73
Q

3 Spirochetes

A

-Borrelia
*burgdorferi: most common, ixodes tick,
lyme/bulls eye/erythema chronicum migran
*recurrentis: relapsing fever

-Treponema pallidum: syphillis, darkfield/silver

-Leptospira interrogans: Weils, zoonotic/urine
-Fletchers medium

74
Q

Mycobacteria

M.TB
-respiratory, tubercles
-cauliflower on LJ
-cording in broth
nonpigmented
-niacin and nitrate pos

M.bovis
-niacin and nitrate neg
-GI infection

M.gordonae
-takes longer 5-10days
-tap water bacilus

M.avium complex
-environmental
-lung disease, AIDs

M.leprae
-Hansen, tissue lepra cells
-armadillo
-acid fast tissue

M.fortuitum/chelonei
-only ones that grow on Mac and grow fastest <7 days

A

Mycobacteria
-increased 5-10% CO2, 35C, 3-6 weeks
-sputum first morning 3 conconsecutive days
-broncial/gastric washings
-urine, tissue
-decontaminate with NaOH and NALC liquefies

DIRECT EXAM
Gram stain:
-structurally gpr but gram invisible since mycolic acid repel CV/iodine
-resist decolorization (acid fast)

3 stains:
-Kinyoun: not heated, uses higher concentration of phenol

-Ziehl Neelsoen: heated so dye can penetrate cell wall; carbol fuschin, acid alcohol, MB

Fite acid fast:
-weaker decolorizer for weak acid fast, nocardia, cryto, cysto/cyclo

-Fluorochrome: more sensitive
*auramine rhodamine, acid alcohol,
potassium permaganate, acridine orange

CULTURE
-gold standard, slow growth
-liquid broth: grows faster; cording M.tb
-2 solid media:
-Middlebrook 7H11: clear
-M=mycobacterium, Middlebrook, microscopically
-Lowenstein Jensen: green
*read uncovered, if growth then read uncovered
-slow or rapid grower
-checked for pigment (scoto in light/dark, Photo in light)

MOLECULAR
-direct nucleic acid amplification

MULTIDRUG RESISTANT TB
-resistant to rifampicin, isoniazid
-susceptibility testing using solid/liquid medium, NAAT TB DNA, LPAs DNA extraction/amplification, NGS/whole genome sequencing extrated TB DNA

75
Q

ACID FAST CONCENTRATION/CULTURE

3-10 ml of sputum is processed and concentrated by centrifugation
* Smears can be made – Directly from processed pellet
* May increase smear sensitivity
* However, may result in less material available for NAA testing & culture – From re-suspended pellet after the addition of buffer
* Re-suspended in ~2 ml buffer; one drop for smear

Prior to heat fixing, smears should be allowed to air dry completely within the biological safety cabinet
* Acceptable methods for heat fixing
* Flame fixing by passing over flame 2–3 times for a few seconds smear side up – Avoid scorching * 2 hrs minimum at 65-75oC on an electric slide warmer
* 15 min at 80oC1 * 5% phenol in 70% EtOH for 5 min2 – also kills AFB

A

AFB tests look for FB bacteria in your sputum. Sputum is a thick mucus that is coughed up from the lungs. It is different from spit or saliva. There are four tests used to identify AFB:

AFB smear. In this test, your sputum sample is “smeared” on a glass slide and looked at under a microscope. It can provide results in 1-2 days. These results can show a possible or likely infection but cannot be used to provide a diagnosis.

AFB culture. In this test, your sample is sent to a lab to allow the cells to grow until there are enough to test. An AFB culture can positively confirm a diagnosis of TB or other mycobacterial infection. But it takes 6-8 weeks to grow enough bacteria to detect an infection.

Molecular test. In this test, your sample is tested using nucleic acid amplification (NAAT) with AFB smear and culture. The NAAT test is done directly on sputum samples and can detect TB bacteria. If there is TB bacteria, the test can also check whether the bacteria are resistant to rifampicin, a common treatment for TB. This test can be done in less than two hours.

Susceptibility test. This test is usually ordered along with an AFB culture. It is used to figure out the most effective antibiotic to treat your infection. Mycobacterium tuberculosis may be resistant to one or more of the medicines that are used to treat TB.

76
Q

Mycobacterium Leprae:
to be diagnostic needs to be from..
lepra cells are….
alternate name…

A

tissue fluid
macrophages w/acid fast bacilli
Hansens disease

77
Q

Tween 80 for what mycobacterium…
neg=
pos=

Mycobacterium test that splits phenolphthalein from triK phenolphthalein-sulfate in 2-3days…

A

Tween 80 M.kansasii
neg=amber
pos=pink

aryl-sulf-atase test

78
Q

Most common bacterial STD, pelvic inflammatory disease

obligate intracellular parasite

A

Chamydia trachomatis

79
Q

Two free living with no cell wall, not bacteria

use antisera and test inhibition of growth

A

Mycoplasma: fried egg, walking/atypical pneumonia

Ureaplasma:

80
Q

Loeffler: C.diph

Lowenstein Jensen: mycobacteria

Regan Lowe: B.pertusus

A

Loeffler: C.diph
-granules, pleomorphism

Lowenstein Jensen: mycobacteria

Regan Lowe: B.pertusus

81
Q

Parasite basics

Divisions:

PROTOZOA subkingdom:
-amebae, flagellates, ciliates, sporozoans, coccidia, microsporidia

NEMATODA phylum: roundworm, unsegmented, complete digestive, males smaller

Platyhelminths phylum: flatworms
-TREMATODES (flukes):
oral/ventral cupshaped sucker, flat leaf, snail/metacercaria
-CESTODES(tapeworms): segmented, flat, proglottids, suckers on scolex

Arthropoda phylum:
-insects, spiders, mites, ticks

Ova
-parasite eggs
Larvae:
- immature form

Cysts:
-inactive stage, formed stool

Troph
-active feeding/reproducing
-loose stools

A

Parasite basics

Collection:
-fecal/intestinal
-vaginal/urethral/urogenital: saline wet swab
-prostatic secretion
-sputum/urine: early morning
-CSF
-tissue/biospy: sterile container
-blood

Stool
-mostly protozoa
-variable schedule so collect over 10 days 1-2days apart
-do before barium sulfate
-no urine/water
–liquid 30min
-soft 1hr
-formed 24hrs

82
Q

Method Detection:

Direct wet mount
-motile protozoan troph, flagellates
-smear made with samples and saline/iodine (iodine for cysts, yellow gold)
-trophs, helminth eggs, larvae, cysts, motility
-Need to id in permanent stain

…

-Saline: motility, low light
-Iodine: cysts, kills troph

…

Preservation/Fix
-intestinal eggs/larvae
-fridge: not for trophs
-formalin/PVA/Schaudinn
-EcoFix, ProtoFix

A

Permanent stain

Trichrome:
-gold std, diff stain
-last part of exam, confirmation
-fixed with PVA before stain
-Crypto/Cyclo/Iso can’t be stained with this
-cytoplasm green or light pink
-background green
-nuclear is red purple

1.)thin film of feces on slide
2.)while wet at Schaudinns 30min RT
3.)Rinse alcohol to remove fixative and add iodine
4.)2 more changes of ethanol slns and rinse with acidified ethanol
5.)Xylene
6.)Mount slides

Modified trichrome/weber green
-microsporidia
-much higher dye

83
Q

Hematoxylin:

-iron, enhance microfilarial sheath or intestinal parasites
-Tompkins miller
-blue gray stained
-uses alcoholic iodine to remove mercuric chloride form Schaudinn’s and PVA-fixed smears
-alcohol washes to remove residual iodine
-a working hematoxylin stain that contains the mordant
-picric acid for destaining; -various concentrations of alcohol and xylene to dehydrate and clear the smear.

A

Wrights, Giesma:
-blood smears
-plasmodium, trypano,microfiliaria, babesia, leish
-Thin: dip in methyl alcohol then 2hrs in giesma
-thick: 50min giesma
-nucleus red, cyto blue

Acid fast: Crytospor, Isospo, Cyclospor
-immunocompromised
-fix with methonol
-Kinyoun carbolfuschin, strain
-ethanol
-sulfuric acid decolorize
-MB
*Crytosporridium is 4-6microns oocysts are pink to deep purple and back is blue
*Confirm with immunoassay antibody technique

84
Q

Method Detection:

Direct Antigen Detection
-O&P lengthy
-antigen detects G.lamblia, Cryptosporidium(water)
-antigen in the stool
-These utilize either an immunofluorescent antibody (IFA) assay or a capture enzyme-linked immunosorbent assay (ELISA) against cyst or trophozoite antigens.
-IIF: monoclonal ab against antigen/cell wall and fluorescein isothiocyanate labeled AHG
-EIA: enzyme conjugated antibody against antigen

Polymerase chain reaction (PCR) techniques may detect giardia in stool samples with parasites concentrations as low as 10 parasites/100 mcL. PCR may also be a valuable tool for screening of water supplies. Real-time PCR has also the advantage of being able to detect both mild and asymptomatic infections.

…

Cellophane tape
-fecal sample doesn’t detect eggs for Enterobius vermicularis
-clear not frosted tape anal opening and placed on slide
-upon waking before bath/bathroom, if neg do on subsequent days
-low/high power, ellipsoid/football shaped, one side flattened

A

Plasmodium
-smears, rbcs, anopheles

…

Tickborne
-Babesiosis: tickborne, maltese cross
-Borrelia burgdorferi: lyme disease, Ixodes tick, detect antibodies serum/csf
-Ehrlichiosis chaffeenis: gn obligate intracelluar parasite in monos, direct visualization of morulae in monos w/Giesma, ab, or PCR

…

Pneumocystis jiroveci
-AIDS
-lung tissue/secretions

85
Q

Fecal concentration

-done in O&P exam, complete ova and parasite
-sedimentation or flotation to enhance detection of smaller #s

-Sedimentation:

uses gravity/centrifugation to recover all protozoa eggs/larvae

*FEA: formalin ethyl acetate:
4 layers, sediment is parasites: top is ethyl acetate, 2nd fecal debris, 3rd formalin, and bottom is sediment/parasites
*detects all but more debris

1.)Fix stool w/formain for 30min: kills, preserves
2.)Strain thru 2 layers of damp gauze into tube
3.)Fill with saline
4.)Centrifuge 10min
5.)Decant and resuspend in saline or formalin
6.)Suspend in formalin and ethyl acetate
7.)Shake vigorously to extract fats, reduce bulk
8.)Centrifuge to get 4 layers
9.)Rim upper debris layer and decant supernatant, and invert once
10.)Add drop of sediment and iodine on slide

A

Fecal Concentration

-Flotation:

separates protozoan cysts and certain helminth eggs; high sg reagent zinc sulfate 1.18 causes parasites to float on surface and debris is on bottom
*cleaner but miss operculated,large eggs

1.)Fix and wash feces as in sedimentation
2.)Wash 1-2x saline centrifuging in between, want 1ml sediment
3.)resuspend and mix in zinc sulfate
4.)Centrifuge and keep vertical while adding more zinc sulfate
5.)Cover slip top to touch meniscus, 10min
6.)Lift cover slip with parasites and add to slide w/iodine, rotate to mix

86
Q

Thick/Thin for blood parasite
-Plasmodium
-Babesia
-Microfilariae (not Onchocerca)
-Trypanosoma
-Leishmania(amastigote in macrophage)

Thick Smear

-Screens
-thick layer of dehemoglinized/lysed RBCs
-more concentrated so more sensitive but can’t see morphology so need to use thin to id

-EDTA whole blood
-slide, applicator
1.)1 drop of blood on slide and spread to make a square thats barely legible thru newsprint
2.)Dry flat
3.)Dip in water quickly and dry
4.)Stain: ph 7.2, Romanowsky stain, Giesma most common, or Wrights 20-30mins, rinse with water
5.)Read under 100x, 100 fields with 20 WBC in each field B4 calling negative
6.)Count 200 wbc in 100 fields

A

Thin Smear

-Do asap after draw
-Smear w/in 1hr

1.)1 drop and smear just like diff and air dry
2.)Romanowsky stain: Giesma, Leishman, ph7.2
-Leishman: stain 1-2 min, water, mix with pipette, leave 4-8min, wash with water, dry
-Chromatin: deep red
-Cytoplasm: blue
3.)Read under 100x:
-Count parasitized RBC as %
-Earliest stage in RBC: ring
-Most common is troph: growing stage
-Schizont: asexual reproduction
-Gametocyte: sexual stage
-P.falciparum: multiple rings, banana gametocytes
-P.vivax: larger and young rbc, shuffner, Fy neg protected
-P.malariae: troph can band, old rbc
-P.ovale: fimbriated rbc

QC:
-one slide read by two
-interlab comparisions

87
Q

Parasite stains used for

-Trichorme:
-Wrights/Giesma
-Acid fast:
-Iron Hematoxylin:
-saline:
-Iodine:

A

Parasite stains:

-Trichorme: nuclear red/purple blue back
-Wrights/Giesma
-Acid fast: spores
-Iron Hematoxylin: microfilarial sheath
-saline: motility
-Iodine: nuclear

88
Q

Preservation:

-2 for cysts,eggs, larvae
-2 for trophs
-1 for trophs, cysts for fresh stool

A

Preservation:

-2 for cysts,eggs, larvae:
-fridge, formalin, MIF

-2 for trophs:
-PVA, SAF

-1 for trophs, cysts for fresh stool:
-Schaudinns

89
Q

Sedimentation:
contains these two, four layers

A

ethylacetate: removes fat/oil but fat/debris stay in separate layer

formalin: preserves

sediment=parasites

90
Q

Concentration:

-FES: ingredients, what 4 layers

-Zinc sulfate flotation: sg

A

Concentration:

FES: ingredients, what 4 layers
-Formalin: preserves
-Ethyl acetate: removes oil/fat
-S: sedimentation (parasites)

Zinc sulfate flotation: sg
-sg 1.18, parasites float to top
-miss operculated large

91
Q

4 Genera of Protozoa

A

Ameba, Flagellates, Ciliates, Sporozoa

92
Q

Trichomonas:
-which one has undulating membrane 1/2 the length, trasmission
-which one has undulating membrane full lenth, transmission

A

vaginalis: 1/2, sexual

hominis: full, feces

93
Q

Trypanosoma:

-West…
-East…
-vector, sickness

A

Both Tse Tse fly, trypomastigote blood

Gambiense:
-West african sleeping sickness

Rhodesiense
-East african sleeping sickness

94
Q

T.cruzi:
-vector
-can be transmitted by…
-disease called…

-amastigote in…
-trypomastigote in…

A

T.cruzi:
-vector: reduvid bug
-can be transmitted by…blood transfusion
-disease called…Chagas

-amastigote in…heart
-trypomastigote in…blood

95
Q

Leismania

-Kala Azar…

-Old…

-New…

A

Leismania

-Kala Azar…dovani

-Old…tropic

-New…braziliensis

96
Q

6 sporozoa genus

A

Plasmodium, Babesia
Crytosporidium, Cyclospora, Isospora
Toxoplasma

97
Q

Isospora, Cyclospora, Crytosporidium and Toxoplasma infective form is ingestion of oocyts

Microsporidia infective form is ingestion of spores/polar tube

A

Isospora, Cyclospora, Cryptosporidium stained with modified acid fast

Microsporidia stained with trichrome

Toxoplasma stained with Giesma

98
Q

Isospora belli is anorexic

Cryptosporidium and Microsporidia have AIDs

Toxoplasma gondii likes cats but kills fetus

Cyclospora just gives you diarrhea

A

…

99
Q

2 Plasmodium that infect young/large RBCs, schuffners dots

-which one has resistance in duffy neg

A

vivax: duffy neg resistance

ovale

100
Q

Plasmodium with Hmg S/G6PD resistance, banana shaped, multiple rings

A

falciparum

101
Q

Sporozoan with four maltese cross ring forms, from deer tick

A

Babesia microti

102
Q

6 Nematodes

A

Enterobius: Pinworm
Trichuris: whipworm
Strongyloides: threadworm
Nectar/ancyclo: hookworms
Ascaris: large intestine roundworm

103
Q

Microfilaria:

Which 2 spread by mosquito, elephantitis, blood:

Which 2 spread by fly
-calabar/eye swelling
-blindness, nodule

A

Microfilaria:

Which 2 spread by mosquito, elephantitis, blood: Wucheria, Brugia

Which 2 spread by fly: loa loa, onchocerca
-loa loa: calabar swelling,eye
-onchocerca: blindness, nodule

104
Q

Sheath, nuclei:

3 have sheaths:
-1 doesn’t have in tip:
-1 has 2:
-1 goes to tip:

1 doesn’t have sheath or nuclei:

A

Sheath, nuclei:

3 have sheaths:
-1 doesn’t have in tip: wucheria
-1 has 2: Brugia
-1 goes to tip: Loa loa

1 doesn’t have sheath or nuclei: onchocerca

105
Q

2 tissue nematodes:
-one with sushi, granuloma:

-one with pigs, encysted larve, muscle biopsy:

A

2 tissue nematodes:
-one with sushi, granuloma:
Aniskasis

-one with pigs, encysted larve, muscle biopsy
Trichinella spiralis

106
Q

Trematodes:

3 intestinal/liver:
1 blood/bladder:
1 lung:

A

Trematodes:

3 intestinal/liver:
-clonorchis: chinese liver
-Fascioloa hepatica: sheep liver
-F.buski: giant GI

1 blood/bladder:
-Shistosoma

1 lung:
Paragonimus westmanii

107
Q

Chinese Liver fluke…

Chinese vase/Lung fluke:

Giant intesting fluke:

Sheep liver fluke:

A

Chinese Liver fluke…Clonorchis

Chinese vase/Lung fluke: Paragonimus

Giant intesting fluke: F.buski

Sheep liver fluke: Fasciola hepatica

108
Q

Blood fluke, Shistosomes spines and location in veins…

-Sml, lateral, intestine veins:

-lrg, lateral, intestine veins:

-lrg terminal, bladder veins:

A

Blood fluke, Shistosomes spines and location in veins…

-Sml, lateral, intestine veins: japonicum

-lrg, lateral, intestine veins: mansoni

-lrg terminal, bladder veins: haematobium

109
Q

5 Cestodes/tapeworms:
ribbon like, proglottids, segmented

A

Tania
Hymenolepsis
D.latum (only operculated)
Echinococcus
D.caninum

110
Q

Names of diseases:

-cutaneous larva migrans/creeping eruption

-larva currens

-erythema chronicum migrans

-visceral larva migrans:

A

Names of diseases:

Strongyloides:
-larva currens

Borellia burgdoferi/Lyme;
-erythema chronicum migrans

Ancyclostoma braziliense
-cutaneous larva migrans/creeping eruption:

toxocara canis/cat:
-visceral larva migrans

111
Q

GPR aerobic spore forming, bamboo pole
string of pearls
nonmotile

A

B,anthracis

112
Q

Gram positve rod, nonmotile
-chinese letters,picket fence
-pallisade, club shaped

A

Corynebacterium diphtheria

113
Q

Phase where bacteria are maturing but not about to divide

A

Lag

114
Q

Plasmodium vector…

Babesia vector

A

Plasm anopheles mosquito

Babesia deer tick

115
Q

H.nana, Diminuta vector

A

Flea, beetle

116
Q

Which fastidious, gram-negative coccobacilli is often associated with trauma after human bites or fights where the skin has been broken by human teeth?

A

Eikenella corrodens is a member of the usual flora of the oral and bowel cavities. It usually occurs in mixed infections as a result of trauma, especially human bites or “clenched fist wounds”. E. corrodens is also seen in persons with poor oral hygiene, causing adult periodontitis and meningitis.

117
Q

In addition to Giardi sp, what other intestinal parasite is clinically important in the United States?

a.
Cryptosporidium sp

b.
Cyclospora sp

c.
Isospora sp

d.
Toxoplasma sp

A

Cryptosporidium sp

The classic ova and parasite diagnostic method is accurate, but technically demanding and time consuming.

Due to the high likelihood of Giardia or Cryptosporidium as the causative parasite, numbers DFA, EIA and ICT assays are available for screening tests.

118
Q

What is an example of streptococcal group test?

a.
Enzyme immunoassays

b.
Immunofluorescent assays

c.
Particle agglutination

d.
Precipitin tests

A

Particle agglutination

Streptococcal grouping tests use a form of particle agglutination in which antibodies are bound to the surface of latex beads which bind to the antigen present in a sample to produce a visible clump.

119
Q

What is a disadvantage of the zinc sulfate flotation method for separating protozoan cysts and helminth eggs in fecal specimens?
Question 177Answer

a.
Some helminth eggs do not concentrate well with this method

b.
The centrifugation times are longer

c.
The preparation is not as clean as the sediment method

d.
There are no disadvantages to the method

A

Some helminth eggs such as operculated eggs and/or very dense eggs such as unfertilized Ascaris eggs do not concentrate well with the flotation method. Their density causes them to sink instead of float.

120
Q

A gram-negative rod > 100,000 col/cc is recovered from a midstream urine. The isolate is swarming over the blood plate, produces hydrogen sulfide on TSIA, and hydrolyzes urea, indole negative and ornithine positive. What is the likely organism identification?
Question 176Answer

a.
Morganella morganii

b.
Proteus mirabilis

c.
Proteus penneri

d.
Proteus vulgaris

A

Proteus mirabilis is a commensal species in the gastrointestinal tract of humans. It is widely disseminated in the environment and recognized as an opportunistic pathogen.

Proteus vulgaris is indole positive and ornithine negative.

Proteus penneri is ornithine negative.

Morganella morganii is indole positive and H2S negative on TSIA.

121
Q

Which organism needs the X and V factors required for fastidious organisms?

A

Haemophilus spp. need the X and V factors required for fastidious organisms.

122
Q

Which organism does not usually cause disease in a host with an intact immune system, but may cause disease in a host with a compromised immune system?

a.
Community acquired infection

b.
Nosocomial infection

c.
Opportunistic pathogen

d.
Spore forming pathogen

A

Opportunistic pathogen is defined as an organism that does not usually cause disease in a person with an intact immune system, but that can cause disease when the host’s immune system has been compromised by disease or other condition that has damaged or changed the immune status of the host.

Community acquired infection
-less than 48hrs since being in healthcare

Nosocomial infection
-while receiving healthcare, 48hours after admission

123
Q

What gram positive cocci are most frequently associated with UTIs in young, sexually active females?

A

Staphylococcus saprophyticus has been associated with UTI’s in young sexually active females. This species adheres more effectively to the epithelial cells lining the urogenital tract than other coagulase-negative staphylococci.

124
Q

What kind of sample can a parasitic infection be diagnosed through the identification of eggs (ova) or larvae?

a.
Blood sample

b.
Fecal sample

c.
Nail scraping

d.
Urine sample

A

Fecal sample

The parasites are residing in the intestinal tract. Stool specimens for ova and parasite studies must be preserved or fixed immediately.

125
Q

What cells do not have a nucleus, or any membrane bound organelles such as mitochondria, and their ribosomes are a smaller size than eukaryotic ribosomes?

A

Prokaryotes:
Bacterial cells are prokaryotes and are less complex in structure than eukaryotic cells. Prokaryotes, from the Greek “before nucleus”. They lack all membrane-bound organelles.
-DNA is in a single chromosome in a nucleoid region
-peptidoglycan cell wall

Eukaryotes:
-true nucleus
-animal, plant, fungal,parasites
-membrane bound nucleus, organelles, cytoskeleton, larger ribosomes
-have mitrochondria

126
Q

How is the use of molecular assays to detect antimicrobial resistance genes limited?

a.
Genes are not responsible for most types of antimicrobial resistance

b.
Genes may be present but not expressed; presence of the gene may not reflect resistance

c.
Large numbers of genes are responsible for all clinically important resistance

d.
Researchers have not identified genes for antimicrobial resistance

A

Genes may be present but not expressed; presence of the gene may not reflect resistance

There is concern about using molecular methods to confirm resistance because of the ability of some bacteria to contain specific resistance genes that may not be expressed. The clinical significance of the presence of the resistance genes in these bacteria is questionable.

127
Q

A 20-year-old male presents to the Emergency room with heavy urethral discharge that had been present for three days. He has been sexually active with several partners and has used no contraceptives. A culture of the discharge grows a gram-negative diplococci with adjacent sides that are flattened and is oxidase positive. What is the likely organism?

A

Neisseria gonorrhoeae is a sexually transmitted, oxidase positive, gram negative diplococci that can cause urethral discharge in males and females.

*no cotton and no calcium alginate swabs, inhibit

128
Q

Thick smear used to..

Thin smear used to…

A

The thick smear is used to screen a larger volume of blood; the thin smear is used to identify the parasite because the parasites are not as distorted as with the thick smear.

129
Q

What is the process of destroying pathogenic organisms, but not necessarily all microorganisms or bacterial spores (endospores)?
Question 191Answer

a.
Antisepsis

b.
Disinfection

c.
Filtration

d.
Sterilization

A

Disinfection:
Disinfection is the process of destroying pathogenic organisms, but not necessarily all microorganisms or bacterial spores (endospores). Physical (moist or dry heat) or chemical (e.g., bleach, phenol, alcohols, aldehydes) means may be used in the process of destroying pathogenic organisms.

Antisepsis:
decreased number of microbes present on skin.
-iodine,alchol,chlorhexidine

Sterilization:
killing/destruction of ALL microbes including spores.
-heat, filtration, oxidation
-dry heat: 171C 1hr
-wet heat/autoclave: steam under pressure 121C 15lb,15min
-filtration: heat sensitive media

130
Q

Cellophane tape preparation

The correct specimen to collect for a pinworm examination is a cellophane tape preparation. This procedure involves swabbing the child’s perianal area with a tongue blade covered with cellophane tape (sticky side out). The sampling should take place first thing in the morning before the child uses the bathroom or bathes.

A

…

131
Q

A 70-year-old female with a previous knee injury has joint replacement surgery. The surgery goes well, and she is discharged from the hospital to a rehabilitation facility to receive post-operative care and physical therapy. One week later the incision becomes inflamed and begins oozing a discharge. A gram stain of the discharge shows many white blood cells and gram-positive cocci in clusters. The isolate recovered is a cream-yellow beta-hemolytic colony that is catalase and coagulase positive. What is the likely organism?

A

Staphylococcus aureus is a gram-positive cocci in clusters that generally yields a cream-yellow beta-hemolytic colony that is catalase and coagulase positive. Streptococcus pyogenes and Enterococcus faecalis are gram positive cocci that are catalase negative. Staphylococcus epidermidis is coagulase negative.

132
Q

Which specific bacteria, parasitic cysts and rare fungal forms contain mycolic acid in their cell wall and retain the basic dye despite acid alcohol rinsing?

A

Acid fast organisms (containing mycolic acid) resist decolorization by acid alcohol destaining and appear the color of the primary stain.

133
Q

What do specimens for parasitic identification include?

a.
Hair, nails, stool, urine, and sputum

b.
Hair, stool, urine, bone marrow, and sputum

c.
Stool, urine, blood, sputum, and tissue biopsies

d.
Tissue biopsies, cartilage, hair, nail, and urine

A

Stool, urine, blood, sputum, tissue biopsies

Specimens for parasite identification come primarily from the intestinal tract, urogenital tract, or prostatic secretions. Specimens can also come from sputum, CSF, or biopsy material from tissues. Malarial parasites can be seen in blood.

Any identification process first depends on correct specimen (stool, urine, blood, sputum, tissue biopsies) collection and adequate fixation.

134
Q

Which four groups can most bacteria be divided into based on gram stain results?

a.
Gram positive bacilli, gram positive cocci, gram positive tetrads, and gram negative bacilli

b.
Gram positive bacilli, gram negative bacilli, gram positive clusters, and gram negative chains

c.
Gram positive bacilli, gram negative bacilli, gram positive cocci, and gram negative cocci

d.
Gram variable bacilli, gram positive diplococci, gram negative rods, and gram positive cocci

A

Gram positive bacilli, gram negative bacilli, gram positive cocci, and gram negative cocci

Tetrads, diplococci, clusters, and chains are descriptions of cellular arrangements not basic shapes. Variable indicates an indistinct staining reaction.

135
Q

When should the concentration of fecal materials be included?

a.
Intestinal formalin

b.
Intestinal ova

c.
Ova and parasite examination

d.
Thick and thin smears

A

Ova and parasite examination

Concentration procedures allow the visualization of small numbers of parasitic organisms that may be missed if only a direct mount is observed.

Various concentration procedures are used, most often sedimentation or floatation techniques.

136
Q

What kind of insects belong to the phylum Arthropoda group?

A

The phylum Arthropoda includes insects, spiders, mites, and ticks. Members of this phylum are a subgroup of parasites and are invertebrates.

137
Q

What gram negative rod is associated with lobar pneumonia in hospitalized patients?

A

Klebsiella pneumoniae is a frequent cause of lower respiratory tract infections among hospitalized patients.

138
Q

What is the etiologic agent in Lyme disease?

A

Diagnosis of Lyme disease depends on finding antibodies in the serum or spinal fluid. The etiologic agent is the bacterium Borrelia burgdorferi.

139
Q

A 10-year-old child presents to his primary care physician with a complaint of severe ear pain. The child had been swimming in the lake several days earlier. The culture from the ear grew a gram-negative rod with a grapelike odor. The colonies were flat and spreading with rough edge and a metallic sheen. It was indole negative and oxidase positive. What is the likely organism?

A

The organism is likely Pseudomonas aeruginosa. Pseudomonas is commonly found in fresh water. P. aeruginosa produce a grape like odor, are oxidase-positive and indole negative.

140
Q

What concentration of oxygen is required for the growth of obligate anaerobic organisms?

A

0% oxygen

Obligate anaerobes must be grown in an atmosphere either devoid of oxygen or with significantly reduced oxygen content.

141
Q

What nutrients in chocolate agar enable organisms to grow on this media but are unable to grow on blood agar?

A

Hemin (X) and nicotinamide adenine dinucleotide (V) factors

Chocolate agar contains lysed red cells which release hemoglobin, hemin (X) and nicotinamide adenine dinucleotide (V) factors.

142
Q

What are the most common causes of bacterial conjunctivitis in children?

a.
Haemophilus influenzae andStreptococcus pneumoniae

b.
Herpes simplex and Varicella zoster

c.
Propionibacterium acnes and Alpha-hemolytic streptococci

d.
Staphylococcus epidermidis and Lactobacillus spp.

A

Haemophilus influenzae and Streptococcus pneumoniae are common causes of bacterial conjunctivitis in children.

Staphylococcus epidermidis, Lactobacillus spp., Propionibacterium acnes and alpha-hemolytic streptococci in small numbers are normal flora of the conjunctival sac.

Herpes simplex and Varicella zoster are viral infections.

143
Q

pshychophiles:
-0-20C
-cold

mesophilic:
-moderate temp 30-37
-most pathogenic bacteria,
Ecoli, Salmonella

thermophilic:
-thrive in high temp, 50-60C
-not pathogenic cuz can’t grow at body temp

hyperthermophilic:
-80-120C

A

Incubators

Most pathogens:
35C host temp, 3-5% CO2, humidified
-Can use candle jar for CO2, only white unsented candles

Fungi:
30C

Campylobacter:
42C

144
Q

Culturing specimens/processing/planting/rejection…

*don’t refrigerate spinal, anaerobic, GC
*no dry swabs

Stool;
-lots of NF
-w/in 2hrs
-Salmonella, shigella
-Campy, Clos diff
-Vibrio, Yersinia
-Sero type S/S, E.coli
-Direct detect/molecular C.difficile etc

Urine:
-bladder normally sterile
-clean catch mid stream
-first morning more concentrated
-process in 1hr or fridge 24hrs
-<10 epis
->100,000 sig if asymptomatic
->1,000 if symptomatic male
-UTI: nitrite, leukocyte esterase, usually Ecoli
-Can be Proetus, Kleb, enterobac,Pseudo
-Gram:centrifuge, 1 drop, 20 fields, 1 organism per field

Catheter tip/intravenous:
Indicate source of sepsis or infected indwelling arterial/venous catheter
-For optimal culture results, transport specimen to the laboratory within 2 hours
-Foley from tubing not bag

Urethral/cervical/gynecological:
-No wood shaft on Chlamydia
-No cotton/calcium alginate on Neisseria
-swab to remove purulent, 2nd swab vigorously swabbing
-cell culture for Chlyamydiae
-NAT hybridization
-Trich wet mt, fungal/yeast KOH
-Bacterial vaginosis: clue cells, gvb

IUD:
-Sterile container
-Store and transport room temperature, preferably within 8 hours of collection.
-Actinomyces sp

A

Culturing specimens/processing/planting/rejection…

Blood:
-sterile collection in nutrient broth media b4 fever
-70% alcohol, iodine, alcohol rinse
-2-3 sets 24hrs
-1:5 to 1:10 dilution blood to broth
-10ml adults, 5ml kids
-can use ARD for antibiotic removal
-6 to 18hrs 35C
-blind subculture: bottles subculture to medium even if growth not visible
-smear made
-checked visually 1xday for a week
-growth: hemolysis, gas bubbles, turbidity, colonies
-Automated Bactec: incubated, rocking, measures CO2 made by bacteria, gas permeable sensors use fluorescence to detect CO2
-S.aureaus, CNS, Strep
-Enterococcus, Enterobac, P.aeruginosa
-Yeast(candida, crypto)

Wound/abcess:
-Eswab: 48 hours, refrigerated or ambient
-Sterile cup: 24 hours refrigerated

Tissue:
-Tissue not in formalin
-Sterile screw capped vial, anaerobic culture transport, bacterial culture transport swab with media, or sterile cup
-RT

CSF:
-sterile tubes
-centrifuged if >1ml, 5/10ml best
-S.pneumo, N.mening, H.flue
-E.coli, L.mono
-Newborns: Grp B, L.mono, E.coli
-Kids/young: S.pneumo, H.influ, N.mening
-Old: S.pneumo, L.mono
-Acute meningitis: >1000 WBC, segs, decreased glucose, increased protein
-Aseptic: 100-500 wbc, lymphs, normal to slight decrease glucose, increase protein

145
Q

Respiratory:
-sputum 1st morning, deep coughing, avoid saliva
-not saliva
-gram stain first
-<10 epi, >25PMNs/lpf best
-contamination if gram stained shows >10 eips/lpf

Lower respiratory
-sputum, bronch wash, aspirate, lavage, brush
-S.pneumo, H.influ
-Mycoplasma pneu, Legionella, M.TB

Throat:
-upper respiratory swab for throat culture group A beta strep
-sore throat/pharyngitis can lead to scarlet fever, rheumatic fever, or acute glomerulonephritis
-2 steriles swabs: one rapid, other sent out if neg

A

More Strep/Throat…

Rapid strep antigen detection
1.)enzyme immunoassay
2.)latex agglutionatin
3.)optical immunoassay:
-Strep A OIA
-light reflectance, appear diff color
-need to extract grp A specific carb antigen with enzyme or acid

Strep A,B,C,F,G serology
-antigen detection:
antigen on cell wall elicits immune; rayon swab of throat; antigen is extracted and is bound to antibody that is bound to flurorescent microparticles; polyclonal rabbit anti Grp A strep antibodies
-Slide: extract strep antigen, put on slide with specific latex coated ab=agg

Culture SBA
-pharygeal area
-beta hemolysis

Other GrpA beta
-PYR +
-Bacitracin 0.04 suspectible

*Mostly Strep A in throat but may see S.pneumo
-alpha heme
-optochin suspectible
-bile soluble

146
Q

Normal Flora/pathogens

Sterile normally:
-CSF, urinary tract, blood, transtrachial aspirate/lung tissue

CSF
-sterile
-H.influ, N.menin, cryto

BLOOD
-sterile
-contaminants: micrococcus, CNS, alpa strep

THROAT, OROPHARNX
-NF: alpha strep, staph, Neiss
-Pathogens: Grp A strep, H.influ, B.pertusis

LOWER RESPIRATORY/SPUTUM
Pathogens:
-H.influ, S.pneumo,Kleb pneumo
-S.aureuas
-Myco TB/pneumo, Legionella, Fungi

Transtrachial Aspirate, lung tisse
-Sterile normally
-Pathogen: anaerobe

A

Normal flora/pathogens

URINARY TRACT
-Sterile
-Pathogens: Ecoli, GNR

GENITAL
-Normal: CNS, dip, lacto
-pathogens: N.gonn, Clam trac, Tric vag, G.vag(clue), Tre pall(darkfield), Grp B strep in pregant, Herpes

EYE:
-NF: alpha, staph, neiss
-Pathogens: Chlamydia, N.gonn
-Mostly in newborns
-pathogen kids, conjunctivitis: H.influ, S.pneumo but other alpha strep and Staph is NF

EAR
-pathogens:
Pseu.aeru:swimmers ear
H.influ/S.pneu: otis media

COLON
-profuse NF
-S/S, campy jejuni, vibrio
-C.difficile, Yersinia enter

GASTRIC
-pathogen: H.pylori: urease

SUPERFICIAL WOUND
-S.aureus, Grp A strep
-Pseu aeru

DEEP WOUND/ABCESS
-anaerobe, aerobe

147
Q

4 BioSAFETY levels

1: well known, do not usually cause disease in healthy, minimal hazard (B.subtilis)

2: routine specimens shigella, HIV, Salmonella

3: not usually encountered routine, Myco TB, arbovirus, arenavirus, mold stages systemic fungi, BSC/protective wear

4: not found in routine, exotic, highly controlled, ebola, arboviruses

*air goes from low risk to high risk

A

…

148
Q

Microscopic morphology;

Pairs and Clusters:
-S.aureus, CNS

Pairs and Chains:
-Group A,B,C,F,G beta hemolytic strep
-Viridians strep
-Enterococci

Lancet shaped, pairs/chains, halo due to capsule
-S.pneumo

A

Macroscopic/Colony morph

Staph aureus
-med to lrge, creamy, beta

CNS
-sml-med, no pigment, gamma

Group A,B,C,F,G beta strep
-mostly small gray white, beta

Strep pneumo
-sml, glistening, mucoid, convex, alpha

Viridans strep
-pinpoint, alpha

Enterococcus
-small, gamma

149
Q

Micro QC:

Media: CLSI Clinical and Lab Stds Institute/NCCLS
-performance testing: checking media with cultures of known stock microbes
-Exempt has QC failure rate of less than 0.5%
-Nonexempt more than 0.5%

DAILY
-Equipment temps
-CO2, reagent pos/neg ctrl

WEEKLY
-Autoclave: spore test
moist heat w/B.stearo
dry heat with B.subtilis
-Antimicrobial tests: subculturing, disk diffusion
-Gram stain ctrl slide Ecoli,S.aureaus

QUARTERLY/4x yr
-microscope

6MTHS
-Centrifuge rpm/speed

Annually
-Balance, BSC

AS NEEDED
-Gas pack for anaerobic methylene blue strip, O2 is blue
-thermometers: ref therm
-Insturment logs

MEDIA
-exempt from retesting if bought from CLSI manufacteer
-retest CHOC for Neis,campy
-Procedure manual reviewed/signed annually, kept 2yrs if obsolete
-compentency on hire/yearly
-Proficiency 80%

A

Culturing:

Primary: dilution streak to get isolated cultures

Subculture: mixed, separate diff types

Pure culture: several colonies from a single colony, a single cell
-grow by binary fission, a cell divides to make 2 daughters

Culture: the growth of microbes on artificial media

Culture medium: mix of nutrients which microbes is grown

Agar: seaweed extract

Microbe req:
-most are faculative anaerobes
-48hrs 35C
-ph 6.5-7.5 neutral
-moisture for metabolic rxns
-3-5% CO2
-store in fridge 4C

150
Q

Autotrophic:
-producers, make own food from raw, photosynthesis

Heterotopic:
-consumer, can’t make own food, humas

Bacteriophage:
-virus that infects and replicates within bacteria and archaea. Phages replicate within the bacterium following the injection of their genome into its cytoplasm.

Cell wall: peptidoglycan, maintain shape, prevent cell lysis

Cell membrane:
-inside cell wall, lipid bylayer around cytoplasm

Cytoplasm:
-the semifluid substance of a cell that is external to the nuclear membrane and internal to the cellular membrane, sometimes described as the nonnuclear content of protoplasm. In eukaryotes (i.e., cells having a nucleus), the cytoplasm contains all of the organelles.

Nucleus:
-contains genome

Capsule:
-slime layer, encloses w/polysaccharide/protein

Spore/endospore:
-hostile environment, resist heat,cold,dry,chemicals; will germinate when safe

Pili:
-attach to host cells surface, hairlike projections, virulence factor

A

Osmosis:
-spontaneous passage and diffusion of water/solvent thru semipermeable membrane; water goes from sln with high concentration to sln low thru cells semipermeable membrane, allowing some things to pass and not others

ambient:
-relates to immediate surrounding of something

aerobic:
-grow w/O2,obligate require

anaerobic:
-doesn’t need O2, obligate can’t grow in O2

faculative aerobe:
-use O2 but can switch

faculative anaerobe:
-can live w/out O2

151
Q

2 Pathogens in eye, ear

A

H.influ, S.pneumo

152
Q

Molecular methods in the microbiology lab include:

Nucleic acid hybridization techniques These techniques use the formation of hydrogen bonds between complementary single-stranded DNA (ssDNA).

Nucleic acid amplification The most common technique used to amplify nucleic acid is polymerase chain reaction (PCR).

In situ hybridization This technique detects DNA or RNA directly in the cell or tissue.

In-solution hybridization This is the most common method used in laboratories.

Real-time PCR This technique uses fluorescent conjugated probes to directly quantify DNA.

Reverse transcription polymerase chain reaction This technique uses the enzyme reverse transcriptase to synthesize DNA from RNA transcripts.

Multiplex polymerase chain reaction This technique involves two or more targets that are detected simultaneously.

Nested polymerase chain reaction This technique uses two consecutive PCR reactions.

Strain typing This technique uses primers or probes to differentiate different strains or species of bacteria.

Plasmid profile analysis This technique uses plasmid genetic DNA instead of chromosomal DNA.

Nucleic acid sequencing This technique works with PCR to determine the order of nucleotides in a given DNA fragment.

Molecular methods in the microbiology lab include:

Nucleic acid hybridization techniques These techniques use the formation of hydrogen bonds between complementary single-stranded DNA (ssDNA).

A

DNA probes, also known as gene probes, are single-stranded DNA or RNA sequences that can be used to identify microorganisms in microbiology. DNA probes are complementary to specific DNA sequences from a particular microbe, and can be used to detect infections caused by microorganisms that are difficult to culture.

Probes are a single-stranded sequence of DNA or RNA that is used to identify specific sequences of DNA or RNA. They are designed as complementary to the part of the genome of interest, so that when the probe and the genome are brought together, the probe will hybridise with the target sequence.

Probes are designed to be detected, and they are often modified to fluoresce or are radioactively labelled so that they can located easily

Probes are used in a number of clinical genomic tests, including fluorescent in situ hybridisation, quantitative PCR (where the amount of PCR product is measured) and microarrays.

153
Q

Clostridium difficile:
antibiotic associated pseudomembranous colitis from A and B toxins

-stool toxin EIA:
detects A and B toxin antigen; chromogenic(blue), enzyme conjugate

-GDH: stool is diluted w/buffer, vortexed, and supernatent tested; Glutamate dehydrogenase made by both toxic and nontoxic C.diff;EIA detects the GDH antigen

-NAAT:
detect C.diff toxin genes tcdA or tcdB

-Toxigenic Culture gold standard, takes several days; cultured then cytotoxic assays test isolates for toxin

A

Helibacter Pylori

-fecal antigen test: monoclonal ab immunochromatography detects pylori ag
-Rapid urease: CLO, urease breaks down urea to ammonia chaning ph and phenol red from yellow to red
-Carbon 13 urea breath:
drink urea labeled with carbon isotope which will be high is urease present; bicard is exhaled as CO2 and measure with scintillation counter or mass spec
-Serology: immunoglobulin G ab against pylori with ELISA

154
Q

Shiga toxin

-STEC: shiga toxin e.coli, 0157:H7
-causes HUS

-enzyme immunoassay to detect Shiga toxin 1 and Shiga toxin 2
-monoclonal ab against St coupled with horseradish peroxidase
-this conjugate binds to Shiga toxin

A

Campylobacter

-surface antigen tested with EIA
-or antigen is tested with lateral flow immunochromatographic
-can use NAAT
-Culture: -microaerophilic, 42C; charcoal based; campy bap

Fecal occult:
-screen, can’t see
-Guaiac chemical from plant coated on card, H2O2 yields blue
-FIT/immunochemical: uses ab to detect and bind hmg/blood and more sensitive to guaiac
-DNA: alterations w/colorectal cancer

155
Q

Ocular micrometer:

-calibrated ruler etched on glass disk
-ocular division/OD: distance between each line
-stage micrometer: each line is 10microns apart
-Do for each objective, new objective, or if moved

A

1.)Ocular micrometer into 10x eye piece
2.)Calibrated stage micrometer slide on stage
3.)Coarse/fine focus engraved lines
4.)Line 0’s up on both ocular and stage micrometers
5.)Find line farthest on right that ocular is superimposed on stage
6.)Count # of OD from 0-superimposed line in stage and then count # OD in ocular
7.)OD stage/OD ocularx1000 microns

156
Q

Gram positive

Catalase: Staph from strep

Coagulase: s.aureus from sapro, epi

Novoniocin: epi from novo cns

Optochin: bile/pneumo from virridans

Bacitracin A: Group A strep from B

Camp/Hipp: Group B from A

PYR: group a from B

Salt: enterococcus from group d strep

A

Gram negative

Oxidase pos: Neisseria, Moraxella from GPC

Sugars: neisseria from moraxella

Dnase: moraxella from neisseria

157
Q

Gram positive Rods aerobic BCLEN

Spores: Bacillus
Nonmotile: B.anthracis

Salicin: L.mono from Corynebacterium

Motility: L.mono from Corynebacterium

Urea: Nocardia

A

GNR with animals:

Bartonella: cat, trench fever

Pasteurella: cat/dog, musty, bipolar

Brucella: goat, cow, milk, undulant fever, bioterrorism

Franciella: rabbit fever, tularemia,bipolar, level 3 pathogen

GNR not animals:
Legionella:AC, Pontiac fever, bcye

Bordetella: whooping cough, Dtap, , Regan lowe, Bordetella Gengou(Mercury droplets)

Gram variable:
G.vaginalis: clue cell, HBT

158
Q

GNR fastidious microaerophillic

Campy: curved, motile, 85N, 10CO2, low O2, jejuni seagulls, chicken, 42C

Heli pylori: stomach, ulcer, urea

A

GNR fastidious GI

Vibrio: choleroea rice water stool, tcbs, comma, string test pos/deoxycholate

Aeromonas: aquatic, not on tcbs

Pleisiomonas: not on tcbs

159
Q

HACEK upper respiratory, mouth
GNCB, oral flora

Haemophilus
Aggregatitor/actinobacillus
Cardio
Eikinella: mouth bite, fist, bleach, pit
Kingella: joint in kids, hemolytic

A

Nonfermentative GNR, need oxygen use OF media

Pseudomoas aeru: oxidase pos, swimmers ear, grape

Acine-tobacter: NF skin

Stenotro maltophilia: cystic fibrosus, ammonia

160
Q

GN Obligate Intracellular bacterial like cell wall, arthropod vector

Tick…
Rickettsia: typhus, pox, spotted, egg yolk sacs, weilfelix/Proteus

Ehrlichia: morulae WBc

Anaplasma: morulae in cells

Mites…
Oriental: scrub typhus, bush

Cattle…
Coxiella: Q fever

Obligate Intracellular parasite…
Chlamydia: trachomatis bacterial STD giesma

No cell wall/smallest free living…
Mycoplasma: walking pneumonia, fried egg, antisera to inhibit growth

Ureaplasma: urogenital, urease, granular Brown A8

A

Enterobacteraceae:
-GNR, glucose, nitrate reduce, oxi neg

-IMViC:
Indole: Kovacs, tryptophan, pink
methyl red: make glucose, drop ph red
Vogas: acetoin, red way to metab glu
Citrate: carbon, pH increase blue
PEE ++- - coli, ed
KES –++ entero, serr

Decarboxylase: LAO

deaminase/PD: PPM

Onpg: slow LF

Nitrate reduction: 1st red is pos, 2nwith zinc color is neg

Urea: PUNCH
Urease: PPM

161
Q

Proteus mirabilis: indole neg

Vulgaris: indole pos

Both swarm

A

1a.)AMT Micro/Para Quick (1 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2024 Bold Learning Solutions. Terms and Conditions