(3) Collection, Handling, Processing of Clinical Mycology Specimens Flashcards

1
Q

Enumerate the Techniques to obtain culture material for slide preparation

A
  1. Tease Mount Method
  2. Cellophane Tape Method
  3. Slide Culture/ Microculture/ Riddel’s Method
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2
Q

Techniques to obtain culture material for slide preparation

  • One of the most common technique done in the laboratory.
  • A dissecting needle, commonly a bent wire, is used to pull apart a fungal colony which is placed on a slide.
  • Usually done when placing an LPCB stain on a culture sample.
A

Tease Mount Method

Go over the procedure

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3
Q

Techniques to obtain culture material for slide preparation

Cellophane tape is used to transfer aerial hyphae from the colony to a microscope slide for examination

A

Cellophane Tape Method

Go over the procedure

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4
Q

Techniques to obtain culture material for slide preparation

Allows for in-situ analysis of fungi with as little disruption on the structures as possible

A

Slide Culture/ Microculture/ Riddel’s Method

go over the procedure

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5
Q

Enumrate the Biochemical Tests

A
  1. Urease Hydrolysis Test
  2. Germ Tube Production
  3. Carbohydrate Assimilation Test
  4. Red Colonial Pigmentation
  5. Woods Lamp Examination
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5
Q

Biochemincal Tests

Tests an organism’s ability to produce exoenzyme, called urease, which hydrolyzes urea to ammonia and carbon dioxide.

A

Urease Hydrolysis Test

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6
Q

Biochemincal Tests

what is the positive reaction for Urease Hydrolysis Test

A

PINK

Urease (+) organisms generate enough ammonia to turn the phenol red indicator in the media into PINK.

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7
Q

Biochemincal Tests

what are the Urease positive fungies

A

Cryptococcus neoformans and
Trichophyton mentagrophytes

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8
Q

Biochemincal Tests

hyphae-like extensions of young yeast cells showing parallel non-septate sides

A

Germ Tube Production

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9
Q

Biochemincal Tests

what are teh fungi positive for Germ tube

A

Candida albicans and Candida dubliniensis

go over the procedure

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10
Q

Biochemincal Tests

  • Determine the ability of a yeast to use a carbohydrate as its sole carbon source.
  • Uses a yeast nitrogen base broth
A

Carbohydrate Assimilation Test

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11
Q

Biochemincal Tests

A (+) result for Carbohydrate ASsimilation test indicated by the growth of yeast means

A

the fungi can use carbohydrate as its carbon source

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12
Q

Biochemincal Tests

  • Use the potato dextrose
    agar
    to enhance the pigment production in some dermatophytes

red pigment is then observed in the sub-surface of the agar surrounding the
fungi

A

Red Colonial Pigmentation

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13
Q

Biochemical Tests

What fungi are red pigment producers

A

Trichophyton rubrum
and Rhodotorula species

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14
Q

Biochemical Test

Diagnostic test in which the skin or hair is examined while exposed to the black light emitted by a wood lamp

A

Woods Lamp Examination

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15
Q

Biochemical Test

Infected skin or hair (+) for wood lamps examination, dermatophytes will fluoresce what color?

A

bright green to yellow-green

The test works for both zoonotic and human infections.

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16
Q

Important in cases of some fungal infections where the fungal polysaccharide or proteins are shed out in body fluids

A

Serology-Antigen Detection Test

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17
Q

Serology- Antigen used

Polysaccharide capsular antigen of Cryptococcus neoformans

A

Glucuronoxylomannan

18
Q

Serology- Antigen used

Mannan Antigens

A

For Candida species

19
Q

Serology- Antigen used

Galactomannan Antigens

A

For Aspergillus species

20
Q

Match the antigen

  1. Galactomannan
  2. Glucuronoxylomannan
  3. Mannan

a. Polysaccharide capsular antigen
of Cryptococcus neoformans
b. Aspergillus species
c. Candida species

A
  1. b
  2. a
  3. c

BACcla

21
Q

Provides information that aids the physician in selecting the appropriate anti-fungal medicine totreat a specific fungal infection.

A

Antifungal Susceptibility Testing (AST)

22
Q

AST results for fungi are easily influenced by methodological factors such as?

A
  • Isolate growth phase and inoculum size
  • Incubation time and temperature
  • Type of media used
  • Method of reading
23
Q

Enumerate the Type of AST

A
  1. Brothe Dilution
  2. Disk Diffusion Method
24
Q

Types of AST

  • Different concentrations of one anti-microbial agent (AMA) against one fungal isolate.
A

Broth Dilution

25
Q

Types of AST

Determines the minimum inhibitory concentration (MIC)—the lowest concentration of AMA that can inhibit the growth of fungi

A

Broth Dilution

26
Q

Type of AST

  • AMA are impregnated onto paper disks.
  • Several AMA with standardized concentrations against one isolate
A

Disk Diffusion Method

27
Q

These standards were formed due to the wide factors that can affect the results of antifungal susceptibility test

A

CLINICAL LABORATORY STANDARDS INSTITUTE (CLSI)
STANDARDS FOR AST

28
Q

AST

what document pertains to reference method for broth dilution antifungal susceptibility testing of YEASTS

A

Document M27-A4

29
Q

AST

Reference method for broth dilution antifungal susceptibility testing of FILAMENTOUS FUNGI

A

Document M38-A3

30
Q

AST

DOCUMENT M38-A3 is of Microdilution method for molds that causes invasive and cutaneous infections including what spp?

A

Aspergillus species, Fusarium species, and Zygomycetes

31
Q

AST

Method for antifungal DISK DIFFUSION susceptibility
testing of yeasts

A

Document M44-A2

32
Q

DOCUMENT M44-A2 are intended only for testing this spp and not for other genera

A

Candida species

33
Q

identify what AST document

Final Inoculum: 0.5-2.5x 103 CFU/mL

Test Medium:Roswell Park Memorial Institute (RPMI) 1640 with 0.2% glucose

Growth Temperature: 35 ‘C

Incubation Time: 24 to 48 hours

Endpoint: 50% inhibition

Reading of Endpoint: Visually

A

DOCUMENT M27-A4

34
Q

identify what AST document

Final Inoculum: 0.4-5 x 104 CFU/mL

Test Medium: Roswell Park Memorial Institute (RPMI) 1640 with 0.2% glucose

Growth Temperature:** 35 ‘C**

Incubation Time: 48 hours

Endpoint: 100% inhibition

Reading of Endpoint: Visually

A

DOCUMENT M38-A3

35
Q

Identify the Antifungal agents

A
  1. Polyene Macrolide Antifungals
  2. Azole Antifungal Drugs
  3. Polyene Macrolide Antifungals
36
Q

Antifungal agents

This group of antifungal agents BINDS TO THE ERGOSTEROL of the fungal cell membrane causing the membrane to weaken and results to fungal cell death

A

Polyene Macrolide Antifungals

37
Q

Antifungal agents

what are the Polyene Macrolide Antifungals

A
  • Amphotericin B
  • Nystatin
  • Griseofulvin
  • 5-Fluorocytosine or Flucytosine
38
Q

Antifungal agents

This group of antifungal agents INHIBITS THE FUNGAL CYTOCHROME P450 ENZYMES

o These enzymes are needed for the synthesis of ergosterol in the cell membrane of fungi

A

Azole Antifungal Drugs

39
Q

Antifungal agents

what are the Azole Antifungal Drugs

A
  • Clotrimazole and Miconazole
  • Fluconazole
  • Ketoconazole
  • Itraconazole
  • Voriconazole
  • Posaconazole
  • Anidulafungin
  • Micafungin
40
Q

Antifungal agents

This group of antifungal agents INHIBITS BETA-(1,3)-D-GLUCAN SYNTHASE which is an important enzyme for the synthesis of the components of fungal cell wall.

A

Polyene Macrolide Antifungals

41
Q

Antifungal agents

What are the polyene macrolide antifungals used

A
  • Caspofungin
  • Selenium Sulfide
  • Potassium Iodide
42
Q

Antifungal agents

INHIBITS SQUALENE EPOXIDASE which is needed for the synthesis of ergosterol in the fungal cell membrane

A

Terbinafine or Lamisil

43
Q

ti

A

te