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Myco-Viro > (2) Collection, Handling, Processing of Clinical Mycology Specimens > Flashcards

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

1
Q

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Enumerate the Specimens to revcover fungi

A
  1. Hair, skin, and nail scrapings
  2. Respiratory tract secretion
  3. Cerebrospinal fluid (csf)
  4. Blood
  5. Vaginal secretion
  6. Urine
  7. Tissue, bone marrow, and sterile body fluid
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2
Q

Specimen to recover fungi

Some of the most common specimens used for diagnostic mycology

A

Hair, skin, and nail scrapings

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3
Q

Specimen to recover fungi

They are commonly used to detect dermatophytes, which are keratin-loving fungi.

A

Hair, skin, and nail scrapings

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4
Q

Specimen to recover fungi

Hair samples are collected through ____, and placed on a glass slide,or if being transferred, place it initially on a paper or envelope

A

plucking

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5
Q

Specimen to recover fungi

Skin scrapings are collected with the use of a sterile blade wherein the site of collection is first cleaned with a ____ to remove normal flora, and then followed by scraping the skin.

A

70% alcohol

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6
Q

Specimen to recover fungi

When the skin has lesions, sample is collected
from?

A

the edge of the lesion where the most active fungi are located

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7
Q

Specimen to recover fungi

how nails are collected?

A

nail clippers

Preferably, the nails with lesions should be collected

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8
Q

Specimen to recover fungi

Used for detecting systemic mycoses which mainly attacks lungs

A

Respiratory tract secretion

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9
Q

Specimen to recover fungi

  • It may involve sputum, bronchial washing, bronchoalveolar lavage, and tracheal aspirates.
  • They are considered as non-sterile and may consist of normal flora apart from the target causative aent.
A

Respiratory tract secretion

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10
Q

Specimen to recover fungi - Respiratory

To prevent interferences from the normal flora, culture media containing inhibitors, specifically?

A

antibiotics

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11
Q

Specimen to recover fungi

  • Collected by physicians through spinal or lumbar tap.
  • It is considered as a sterile specimen, and no normal flora are present within the samples. (culture without inhibitors can be used)
A

Cerebrospinal fluid (CSF)

used

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12
Q

Specimen to recover fungi

what portion of CSF samples are tested?

A

Sediment part

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13
Q

Specimen to recover fungi

how sediment part of CSF isolated?

A
  • First Method: membrane filter is attached in the syringe and is used to separate the sediment
  • Second Method: centrifugation

the liquid portion should not be thrown, and should be kept at room temperature

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14
Q

Specimen to recover fungi

Also mainly used to detect systemic fungi

A

BLOOD

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15
Q

Specimen to recover fungi

how is blood samples examined?

A

automated machines

most common automated machines used in the laboratory are BACTEC, VERSATEK, and Bact-ALERT - mainly used to detect yeast

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16
Q

Specimen to recover fungi

  • Commonly used in detecting infections mainly caused by the Candida species.
  • Samples are collected through a cotton swab and are processed immediately as it can dry out which disintegrates the fungi.

ASAP Procesing

A

Vaginal Secretion

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17
Q

Specimen to recover fungi

  • Processed immediately, within 24 hours, upon receipt.
  • The sediment is also the portion examined and may be collected through centrifugation
A

Urine

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18
Q

Specimen to recover fungi

  • Processed immediately upon receipt.
  • Time of collection is noted since the optimum time of recovery of fungi is within an hour after collection
A

Tissue, Bone marrow, and sterile body fluids

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19
Q

Specimen to recover fungi

Samples should be submitted in the laboratory and
processed as soon as possible.

BUT WHAT IF THERE IS A DELAY? what can u do

A

Refrigeration for short time, max of 24 hours
- EXCEPT for five specimens— hair, nail, skin, blood, and CSF. Stored at RT or incubaterd at 30 ‘C
- When storing exceeds 24 hours, samples should be recollected.

quit yo job

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20
Q

Enumerate the methodologies for fungal infection

A
  1. Microscopic Examination (Direct and Staining)
  2. Fungal Culture
  3. Biochemical Tests
  4. Serology-Antigen Detection Tests
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21
Q
  • One of the routine analysis in the laboratory
  • Provides rapid diagnosis of some fungal infections. HOWEVER just the presence or absence of fungi
  • Look for fungal structures
A

Microscopic Examination

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22
Q

T or F

  1. Direct examination require STAINS to be added
  2. Samples are examined in HIGH LIGHT
A

Both F

  1. Does not require stain - COLORLESS
  2. Examined under low light
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23
Q

Direct Examination

  • Examination is done in a natural environment.
  • Good for examining YEASTS.
  • Only incorporates saline.
A

Saline Wet Mount

structure.

Report as either “positive” or “negative” for fungal

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24
Q

Direct Examination

Commonly done on skin scrapings, hair, nails, sputum, and
vaginal secretions.

A

Potassium Hydroxide (KOH) Preparation

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25
Q

Direct Examination

What is the function of KOH preparation?

A

softening, digesting, and clearing cellular debris, such as tissue cells and mucous, to prevent
interferences

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26
Q

Direct examination

what percentage of KOH are used for routine and hard exterior

A
  1. 10% KOH
  2. 20% KOH
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27
Q

Direct Examination

KOH preparation shoud be place in a petri dish together with?

A

Damp piece of filter paper
to prevent speciment from drying out

Report as either “positive” or “negative” for fungal

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28
Q

Direct Examination

Aids in the visualization of diagnostic features of fungi, especially those that are dimorphic

A

Stains

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29
Q

Enumerate all the stains

A
  1. Lactophenol Cotton Blue (LPCB)
  2. Calcofluor White
  3. India ink
  4. Giemsa or Wright Stain
  5. Gram Stain for Yeasts
  6. Periodic Acid Schiff (PAS)
  7. Gomori Methenamine Silver Stain
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30
Q

Stains

It is one of the most common stains used in the laboratory during routine tests

A

Lactophenol Cotton Blue (LPCB)

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31
Q

Stains

Enumeate the component of LPCB

A
  • Lactic Acid
  • Phenol
  • Cotton Blue (aman stain)
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32
Q

Stains - LPCB

used as a preservative for LPCB

A

Lactic acid

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33
Q

Stains

Killing or removal of non-fungle element for LPCB

A

Phenol

34
Q

Stains

the actual stain for LPCB

A

Cotton blue or Aman stain

35
Q

Stains

  • Fluorescent dye that binds with the chitin in the fungal cell wall.
  • Requires a fluorescence microscope
A

Calcofluor White

36
Q

Stains

what is the appearance of Calcfluor white stained fungi under a micrscope

A

appear white to blue to green fluorescence

37
Q

Stains

what can be mixed with Calcofluor white for rapid diagnosis of fungal structure

A

KOH

38
Q

stains

A negative or an indirect stain used
for the detection of the CAPSULE of
Cryptococcus neoformans in CSF
specimen

A

India ink

39
Q

Stains

what is the causative agent of meningitis, and give the stain for the detection

A
  • Cryptococcus neoformans
  • India ink
40
Q

Stains

what will be the appearance of capsule with india ink stains

A

Appear as clear halo against a black background

41
Q

Stains

Used for the examination of bone marrow or peripheral blood sample

A

Giemsa or Wright stain

42
Q

Stain

what does the Giemsa stain detect?

A

Histoplasma capsulatum—a type of systemic fungi

43
Q

Stain

  • Routine stain for the detection of bacteria but can also detect
    fungi if present in the specimen.
  • Yeast stains as gram (+) which is indicated by a purple
    color.
A

Gram Stain for Yeasts

44
Q

Stain

  • Stains the polysaccharide in the cell walls of fungi
  • Fungi stain pink-red with blue nuclei
  • Most commonly used for tissue samples collected through biopsy
A

Periodic Acid Schiff (PAS)

45
Q

Stain

Silver nitrate outlines fungi in black due to the silver precipitating on the fungal cell wall.

A

Gomori Methenamine Silver Stain

46
Q

Stain

what are the staining color of the internal parts of the hypae and background for Gomori stain

A
  • Deep rose to black
  • Light green
47
Q

Considered to be the GOLD STANDARD for the diagnosis of fungal infection

it detects specific GENUS and SPP

A

Fungal Culture

48
Q

TOF - Fungal Culture

Advantage of yielding the SPECIFIC ETIOLOGICAL AGENT through fungal culture—in vitro using artificial media

A

T

49
Q

Fungal Culture Considerations

  1. Incubation temp (most fungal)
  2. Growth for most fungi
  3. Incubation temp for Dimorphic
  4. Growth for rapid growers
  5. Cultures maintenance

a. 37 ‘C
b. 25 ‘C to 30 ‘C.
c. 1 to 2 weeks
d. 21 to 30 days
e. high-humidity environment

A
  1. B
  2. D
  3. A
  4. C
  5. E
50
Q

Culture media

what media are used for anti-fungal agent which removes fungal
contaminants

A

Media with and without CYCLOHEXAMIDE

51
Q

Culture media

What media are used for required for samples with normal flora or
non-sterile samples

A

with and without an ANTIBACTERIAL AGENT

52
Q

Culture media

Medium may be place in a??????

A

plate or tube

plated media is preffered offers a larger surface for growth

53
Q

What are the types of primary culture media

A
  1. Primary or General Purpose Media
  2. Differential Test Medium
54
Q

Contains nutrients necessary for the growth of a wide variety of fungi—making it a non-selective medium

A

Primary or General Purpose Media

55
Q

what is the most commonly used general media

A
  1. SDA - mold culture (It can also inhibit the growth of bacteria due to its acidic pH)
  2. BHIA - for pathogenic fungi, specifically, yeast.
56
Q

Identify what Primary Media

  • General media, recommended for
    most studies.
  • Has an acidic pH at 5.6 that inhibits most bacteria and supports the growth of majority of fungi
A

Sabouraud’s
Dextrose
Agar (SDA)

57
Q

Identify what Primary Media

  • Primary recovery of saprobic and pathogenic fungi.
  • Provides a rich medium for bacteria, yeast, and pathogenic fungi.
A

Brain Heart
Infusion Agar
(BHIA)

58
Q

Identify what Primary Media

  • Primary recovery of pathogenic fungi exclusive of dermatophytes.
  • The agar provides a rich medium for yeast and pathogenic fungi.
A

BHIA with Antibiotics

59
Q

Identify what Primary Media

  • Recovery of fungi from blood
  • Enhances the recovery of yeasts in blood
A

BHI Biphasic Blood Culture
Bottle

60
Q

Identify what Primary Media

  • Isolation and presumptive identification of yeast and filamentous fungi.
  • contains substrates that react with
    enzymes produced by different organisms that result in the production of characteristic color changes.
A

Chromogenic
Agar

61
Q

Identify what Primary Media

  • Primary recovery of dermatophytes; recommended as screening medium
  • Dermatophytes produce alkaline metabolites, which raise the pH and change the medium from red to yellow
A

Dermatophyte Test Medium

62
Q

Identify what Primary Media

  • Primary recovery of pathogenic fungi exclusive of dermatophytes
  • Examine plates for growth. Chloramphenicol inhibits bacterial growth.
A

Inhibitory Mold Agar

63
Q

Identify what Primary Media

  • Primary recovery of saprobic and pathogenic fungi
  • Growth is enhanced by a pH alkaline reaction of fungus. Chloramphenicol and antibiotics inhibit the growth of bacteria and
    nonpathogenic fungi
A

Potato Flake
Agar

64
Q

Identify what Primary Media

  • Primary recovery of dermatophytes
  • Inhibits bacteria and saprophytic fungi
A

Mycosel

65
Q

Identify what Primary Media

  • Primary recovery of saprobic and pathogenic fungi
  • Isolates and enhances growth of pathogenic fungi
A

Sabouraud’s Dextrose with Brain Heart Infusion (SABHI)

66
Q

Identify what Primary Media

  • Primary recovery of pathogenic fungi exclusive of dermatophytes
  • Enhances the recovery of Blastomyces and Histoplasma capsulatum from contaminated
    specimens
A

Yeast Extract Phosphate Agar

67
Q

This medium used to enhance pigment development, conidia production, and mold-to-yeast phase transition of dimorphic fungi

A

Differential Test Medium

68
Q

It is able to differentiate species as it is a selective media which allows the growth of a specific fungi, or a common group of fungi

A

Differential Test Medium

69
Q

Identify what Differential Test Medium

Detection of ascospores in
ascosporogenous yeasts

A

Ascospore Agar

70
Q

Identify what Differential Test Medium

Identification of Cryptococcus, Trichosporon, and Rhodotorula species

A

Christensen’s Urea
Agar

71
Q

Identify what Differential Test Medium

Identification of Candida albicans by chlamydospore production; identification of C. albicans by microscopic morphology

A

Cornmeal Agar with Tween 80 and Trypan Blue

72
Q

Identify what Differential Test Medium

Conversion of the dimorphic
fungus Blastomyces speciesfrom
mold to yeast form

A

Cottonseed Conversion Agar

73
Q

Identify what Differential Test Medium

Differential identification of
Aspergillus species

A

Czapek’s Agar

74
Q

Identify what Differential Test Medium

Identification of Cryptococcus neoformans and Cryptococcus gattii

A

Niger Seed Agar
(Birdseed Agar)

75
Q

Identify what Differential Test Medium

Detection of nitrate reduction to confirm Cryptococcus species

A

Nitrate Reduction Medium

76
Q

Identify what Differential Test Medium

Demonstration of pigment
production by Trichophyton rubrum;
preparation of microslide vultures and sporulation of dermatophytes

A

Potato Dextrose Agar

77
Q

Identify what Differential Test Medium

Identification of Microsporum audouinii

A

Rice Medium

78
Q

Identify what Differential Test Medium

Identification of Trichophyton species

A

Trichophyton Agars 1-7

79
Q

Identify what Differential Test Medium

Detection of Cryptococcus species; differentiate Trichophyton mentagrophytes from Trichophyton rubrum; detection of Trichosporon species

A

Urea Agar

80
Q

Identify what Differential Test Medium

Identification of yeasts by
determining fermentation

A

Yeast Fermentation Broth

81
Q

Identify what Differential Test Medium

Identification of yeasts by
determining carbohydrate assimilation

A

Yeast Nitrogen-base Agar

82
Q
A

Myco-Viro (25 decks)

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