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Clinical Bacteriology (Prelim) > (Laboratory) Gram Staining and Acid-fast Staining > Flashcards

(Laboratory) Gram Staining and Acid-fast Staining Flashcards

1
Q

Gram Stain is named after the Danish Bacteriologist who originally devised it in 1882 (published 1884), _____

A

Hans Christian Gram

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2
Q

Gram Stain is named after the Danish bacteriologist who originally devised it in _____ (published _____), Hans Christian Gram

A

1882
1884

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3
Q

The primary stain of the Gram’s method

A

Crystal Violet

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4
Q

Crystal Violet is sometimes substituted with _____

A

Methylene Blue

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5
Q

Gram-positive organisms appear _____ under a microscope

A

Violet/Blue

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6
Q

Gram-negative organisms appear _____ under a microscope

A

Pink/Red

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7
Q

Principle of Gram Staining

A

Gram Staining is based on the ability of bacteria cell walls to retain the crystal violet dye during solvent treatment.

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8
Q

Mordant in Gram Staining Method

A

Gram’s Iodine

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9
Q

Reagent used in Gram Staining to enhance the staining capability of the primary dye

A

Gram’s Iodine

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10
Q

Acts as decolorizer; removes the primary dye in Gram Staining

A

Acetone Alcohol

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11
Q

Secondary dye/Counterstain; to give stain on bacteria that has been wash off upon applying decolorizer in Gram Staining

A

Safranin

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12
Q

_____ stains many Gram-negative bacteria more intensely than does Safranin, making them easier to see.

A

Basic Fuchsin

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13
Q

Some bacteria which are poorly stained by Safranin, such as _____ and _____ and some anaerobic bacteria, are readily stained by Basic Fuchsin, but not Safranin.

A

Haemophilus spp.
Legionella spp.

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14
Q

Based on the Lecture Video, list down the Equipment used in Gram Staining:

A

Bunsen Burner/Alcohol Lamp
Microscope Slide
Water

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15
Q

Based on the Lecture Video, list down the Reagents used in Gram Staining (on chronological order):

A

Crystal Violet
Gram’s Iodine Solution
Acetone or Ethanol
Basic Fuchsin Solution

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16
Q

In Gram Staining, we smear a very thin layer onto the slide, using _____

A

Wooden Stick

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17
Q

True or False

We use cotton swab in applying or layering the specimen onto the slide in Gram Staining.

A

False

We do not use cotton swab as the cotton fibers may appear as artifacts.

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18
Q

In Gram Staining, spread the culture with an inoculation loop to an even thin film over a circle of _____ in diameter

A

1.5 cm

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19
Q

Time for Crystal Violet application in Gram Staining

A

60 Seconds or 1 Minute

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20
Q

Time for Gram’s Iodine application in Gram Staining

A

120 Seconds or 2 Minutes

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21
Q

Time for Acid Alcohol application in Gram Staining

A

30 Seconds

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22
Q

Time for Safranin application in Gram Staining

A

60 Seconds or 1 Minute

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23
Q

The characteristic feature of the genus Mycobacteria.

A

Acid-fastness

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24
Q

Aside from Mycobacteria, acid-fastness can also be seen (although only partial/weak) on certain bacteria namely:

A

Nocardia spp.
Gordona
Rhodococcus
Tsukamurella
Legionella micdadei

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25
Q

A type of lipid and repels water

A

Mycolic Acid

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26
Q

Preferred to do in Patient Preparation for AFB Stain

A

Gargle with lukewarm distilled water only

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27
Q

Preferred not to do in Patient Preparation for AFB Stain

A
  • Do not gargle with mouthwash
  • Brushing your teeth is prohibited
  • Do not eat
  • Do not drink
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28
Q

Preferred specimen for AFB Staining

A

Early morning sputum

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29
Q

In AFB Staining, if the patient is ambulatory, use _____

A

Aerosol-induced collection

30
Q

We only do the aerosol-induced collection if the patient is _____

A

Ambulatory

31
Q

Collect at least _____ or _____ for DSSM

A

1 Teaspoonful or 5-10 mL

32
Q

Collect at least 1 teaspoonful or 5-10 mL for _____

A

DSSM (Direct Sputum Smear Microscopy)

33
Q

For Xpert MTB/RIF, sputum sample should _____

A

Not be less than one (1) mL

34
Q

For _____, sputum sample should not be less than one (1) mL

A

Xpert MTB/RIF

35
Q

The etiologic agent of tuberculosis

A

Mycobacterium Tuberculosis

36
Q

The most commonly encountered acid-fast bacteria, typified by Mycobacterium tuberculosis, the etiologic agent of tuberculosis.

A

Mycobacteria

37
Q

The Acid-fast stained smear is red at _____

A

1000x magnification

38
Q

If Malachite Green has been used as a counter stain, the background, non-acid fast organisms, debris, and other host cells will be stained _____

A

Green

39
Q

If _____ has been used as a counter stain, the background, non-acid fast organisms, debris, and other host cells will be stained green.

A

Malachite Green

40
Q

If Methylene Blue has been used as a counter stain, the background, non-acid fast organisms, debris, and other host cells will be stained _____

A

Blue

41
Q

If _____ has been used as a counter stain, the background, non-acid fast organisms, debris, and other host cells will be stained blue

A

Methylene Blue

42
Q

Also known as “Hot Method” in AFB Staining

A

Ziehl-Neelsen Method

43
Q

A method in AFB Stain that uses heat as a mordant

A

Ziehl-Neelsen Method

44
Q

Ziehl-Neelsen Method uses _____ as a mordant

A

Heat

45
Q

Ziehl-Neelsen Method uses heat as a _____

A

Mordant

46
Q

Also known as the “Cold Method” in AFB Stain

A

Kinyoun Acid-fast Method

47
Q

What is added in the primary stain for Kinyoun Acid-fast Method?

A

Phenol

48
Q

In the Lecture Video, list down the Reagents used for AFB Staining (Ziehl-Neelsen Method) in chronological order:

A

Aqueous Carbol Fuchsin
Acetone Alcohol
Methylene Blue

49
Q

In the Lecture Video, list down the Equipment used for AFB Staining (Ziehl-Neelsen Method):

A

Bunsen Burner/Alcohol Lamp
Alcohol-cleaned Microscope Slide
Applicator Stick
Tripod
Aluminum Foil
Water

50
Q

Preferred size of sputum specimen in smear

A

Thumb-shaped Size

51
Q

Time for applying Carbol Fuchsin Solution in AFB Stain

A

5 minutes with heating in between

52
Q

Time for applying Acid Alcohol in AFB Stain

A

30 Seconds

53
Q

Time for applying Methylene Blue in AFB Stain

A

60 Seconds or 1 Minute

54
Q

In AFB reading interpretation, reading the slide from left to right is equivalent to _____

A

150 Fields

55
Q

In AFB reading interpretation, reading the slide from left to right, and the going down to read the slide from right to left is equivalent to _____

A

300 Fields

56
Q

Acid-fast organisms appear _____ under a microscope

A

Red Color

57
Q

Non-acid-fast organisms appear _____ under a microscope

A

Blue Color

58
Q

Causes of False-positive Acid-fast Smear:

A
  1. Changes in the cell wall.
  2. Insufficient decolorization.
  3. Laboratory contamination.
  4. Delayed processing and overgrowth of other bacteria.
59
Q

Causes of False-negative Acid-fast Smear:

A
  1. Overzealous decontamination.
  2. Loss from concentration techniques.
  3. Organism obscured by a very thick smear.
  4. Over decolorization.
  5. Poor counterstaining.
  6. Lack observer proficiency in reading sputum smears.
60
Q

No AFB observed in 300 Oil Immersion Field is equivalent to _____ in WHO Scale for AFB

A

0

61
Q

1-9 AFB/100 OIF is equivalent to _____ in WHO Scale for AFB

A

+n

62
Q

10-99 AFB/100 OIF is equivalent to _____ in WHO Scale for AFB

A

1+

63
Q

1-10 AFB/OIF in at least 50 fields is equivalent to _____ in WHO Scale for AFB

A

2+

64
Q

> 10 AFB/OIF in at least 20 fields is equivalent to _____ in WHO Scale for AFB

A

3+

65
Q

A 0 in WHO Scale for AFB means _____

A

No AFB observed in 300 Oil Immersion Field

66
Q

An n+ in WHO Scale for AFB means _____

A

1-9 AFB/100 OIF

67
Q

A 1+ in WHO Scale for AFB means _____

A

10-99 AFB/100 OIF

68
Q

A 2+ in WHO Scale for AFB means _____

A

1-10 AFB/OIF in at least 50 fields

69
Q

Aside from Mycobacteria, two other Gram-positive genera, namely _____ and _____, may stain Acid-fast by modified method.

A

Nocardia
Rhodococcus

70
Q

A 3+ in WHO Scale for AFB means

A

> 10 AFB/OIF in at least 20 fields

Clinical Bacteriology (Prelim) (17 decks)

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