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Seminar 1 > (P) Automated ID & AST > Flashcards

(P) Automated ID & AST Flashcards

1
Q
  • most common automated methods of Clinical Bacteriology
  • manufactured by Thermo Fischer Scientific
  • A partner in the battle against resistance
A

Sensititre system

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2
Q

The technique used in Sensititre:

2

A

MD (Broth Micro Dilution)

utilizes CAMHB (Cation-Adjusted Mueller Hinton Broth)

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3
Q
  • It is a workflow standard that is applicable for most of the automated ID and AST found in the microbiology department
  • flow on how to prepare the bacterial isolates for automated
    ID and AST
A

SENSITITRE WORKFLOW

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4
Q

Correct the Sequence

  1. Measure McFarland suspension with Nephelometer
  2. Prepare a McFarland suspension
  3. Select colonies
  4. Read and report results
  5. Load the cards in ARIS 2X, automatically incubate, then
    read plates
  6. Automatically inoculate panels using AIM
  7. Transfer bacterial suspension to the appropriate
    inoculation medium
A

321 76 54

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5
Q

Select Colonies

Select how many colonies from the fresh primary agar?

A

3-5

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6
Q

Select Colonies

Colony for gram (+) bacteria

A

BAP

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7
Q

Select Colonies

Colony for gram (-) bacteria

A

McConkey Agar

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8
Q

Prepare a McFarland suspension

  1. Emulsify the isolate into a ???
  2. adjust it to ??? McFarland standard
A
  1. sterile water
  2. 0.5
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9
Q

Measure McFarland suspension with Nephelometer

  1. This is used to precisely and accurately adjust it to McFarland standard
  2. Homogenize the inoculum using a?
A
  1. Nephelometer
  2. Vortex
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10
Q

Automatically inoculate panels using?

A

AIM

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11
Q

Where do you load the cards to automatically incubate, then
read plates?

A

ARIS 2X

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12
Q

Load the cards in ARIS 2X, automatically incubate, then

read plates

TOF. After dosing plate with inoculum or broth, the plate
is loaded into an OFFLINE incubator.

A

T

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13
Q

Read and report results

As plates are incubated overnight, results can be read and reported. It can be read manually or by using the?

A

Sensititre Optiread

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14
Q
  • Automatic turbidity measurement
  • Standardize all organism suspensions
  • Precise measurement for best results
A

Sensititre nephelometer

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15
Q

SENSITITRE NEPHELOMETER

  • provides an optical density comparable to the density of a bacterial suspension

Anong standard

A

0.5 McFarland standard

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16
Q

SENSITITRE NEPHELOMETER

When using a Sensititre nephelometer, a bacterial suspension is typically standardized to a density equivalent to a 0.5 McFarland standard which roughly corresponds to a concentration of around?

A

1.5 x 10^8 colony-forming units (CFU) per milliliter (mL).

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17
Q

SENSITITRE NEPHELOMETER

TOF. One must be careful in preparing the suspension because
lower inoculum concentrations may lead to false resistant
results with some antimicrobial agents

A

F (higher)

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18
Q

What guidelines stated that there should be a specific broth to be used depending on which organism or isolates would undergo susceptibility testing

A

CLSI guidelines

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19
Q

SENSITITRE PLATE: NON-FASTIDIOUS ORGANISMS

enterobacteriaccea
* Disk diffusion:
* Broth dillution:

A
  • Mueller Hinton Agar
  • Cation-Adjusted Mueller Hinton Broth
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20
Q

SENSITITRE PLATE: FASTIDIOUS ORGANISMS

Streptococcus pneumoniae
* Disk diffusion:
* Incubation of Disk:

A
  • MHA with 5% Sheep Blood Agar
  • 5% CO2 for 20 to 24 hours
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21
Q

SENSITITRE PLATE: FASTIDIOUS ORGANISMS

Haemophilus influenzae

  • Disk diffusion:
  • Broth Dilution:
  • Incubation of Disk:
  • Dilution methods:
A
  • Haemophilus Test Medium (HTM)
  • HTM broth
  • 5% CO2 for 16 to 18 hours
  • Ambient air must be sufficient and must be incubated for 20 to 24 hours
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22
Q

SENSITITRE PLATE: ANAEROBIC ORGANISMS

  • Medium:
  • Incubated:
A
  • Brucella agar with hemin
  • anaerobically for 46 to 48 hours
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23
Q

GRAM-POSITIVE SENSITITRE PLATE

Each GPID has how many dried biochemical tests?

A

32

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24
Q

GRAM-POSITIVE SENSITITRE PLATE

This includes 2 components

A

sugars (glucose, glycerol, maltose) and fluorogenic
substrate tests

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25
Q
  • there is doubling dilution or concentration of anibiotics.
  • The combination of antibiotics and concentrations are based on the CLSI guidelines.
A

GRAM-NEGATIVE SENSITITRE PLATE

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26
Q
  • last drug of choice for multi-drug resistant organisims (MDROS)
A

CUSTOMIZED PLATE WITH COLISTIN

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27
Q

CUSTOMIZED PLATE WITH COLISTIN

TOF. It is impossible to maximize the plate because there are only
five (5) antibiotics.

A

F (possible)

28
Q

CUSTOMIZED PLATE WITH COLISTIN

To maximize plate, run how many isolates at the same time?

29
Q
  • consists of anidulafungin, amphotericin B, micafungin, caspofungin, posaconazole, etc.
A

YEAST & FILAMENTOUS FUNGI AST

30
Q

YEAST & FILAMENTOUS FUNGI AST

at 24 hours, the endpoints are typically easily defined and the MIC is read at the lowest drug
concentration that prevents any discernible color change from
red to blue

A

Ampothericin B

31
Q

YEAST & FILAMENTOUS FUNGI AST

  • trailing endpoint is experienced
  • occurs when a slight color change persists and is often identical in several concentrations
  • The MIC should be read as the first well showing a less intense color change compared to the positive growth control well
  • Any change of color should be the MIC
32
Q

Anti-fungal or anti-TB drugs coming from the CDC recommendation

A

MYCOBACTERIUM TB AST

33
Q

IDENTIFICATION PLATE

  • ID panels can test # isolates
  • Consists of # fluorescents or biochemical tests
A
  • 3 isolates
  • 32
34
Q

IDENTIFICATION PLATE

TOF. ID panels can run 3 isolates at the same time gram stain for gram positive or negative

35
Q

SUSCEPTIBILITY

  • How many isolates per plate?
  • Can report how many antibiotics?
A
  • 1 plate per 1 isolate
  • 15-20 antibiotics
36
Q
  • Automatic plate inoculator
  • Eliminate skipped wells
  • to dispense the content of the broth or isolates to the specific wells
  • no aerosols which will mitigate exposure risk (benefit)
A

Automatic Inoculating Machine (AIM)

37
Q

AIM has a component that decrease contamination

A

Doseheads – decrease contamination

38
Q
  • May be an alternative to AIM – for those who cannot afford to buy a automatic inoculating machine
  • Uses a multichannel pipette to aspirate and dispense the sample (broth or isolates)
A

Sentititre pipette

39
Q

COMMONLY USED SUSCEPTIBILITY TESTING:

4

A
  1. MACROBROTH DILUTION
  2. MICROBROTH DILUTION
  3. GRADIENT TESTS
  4. DISK DIFFUSION
40
Q

COMMONLY USED SUSCEPTIBILITY TESTING:

  • The test tube is utilized and incorporates increasing two-fold dilutions of antibiotics (i.e. 1, 2, 3, 4 ug/mg) in test tubes
  • Minimum inhibitory concentration (MIC) is generated to determine the susceptibility of an organism to a drug.
  • It is the lowest concentration of an antimicrobial that results in the inhibition of the visible growth of a microorganism.
A

MACROBROTH DILUTION

41
Q

COMMONLY USED SUSCEPTIBILITY TESTING:

TOF. The more “potent” the antibiotic, the less is needed to kill the bacteria, and the MIC is lower.

MACROBROTH DILUTION

42
Q

COMMONLY USED SUSCEPTIBILITY TESTING:

  • Miniaturized version of broth macro dilution
  • There is a freeze-dried antibiotic incorporated in individual wells.
  • Several antibiotics can be tested in a range of two-fold dilutions on a single device
  • An MIC is generated to determine the susceptibility of an organism to a drug
A

MICROBROTH DILUTION

43
Q

COMMONLY USED SUSCEPTIBILITY TESTING:

A small disposable plastic called a microdilution tray contains how many wells?

44
Q

COMMONLY USED SUSCEPTIBILITY TESTING:

  • Includes Etest and MTS
  • Employs thin plastic strips that are impregnated on the underside with a dried antibiotic concentration gradient and are marked on the upper surface with a concentration scale
  • plastic is impregnated with dried antibiotic organisms.
  • a two-fold increase in antibiotic concentration that is marked on the upper surface of the concentration scale
A

GRADIENT TESTS

45
Q

COMMONLY USED SUSCEPTIBILITY TESTING:

TOF. After overnight incubation, tests are read by the intersection of the UPPER part of the ellipse-shaped growth inhibition area with
the test strip.

Gradient test:

46
Q

COMMONLY USED SUSCEPTIBILITY TESTING:

  • called “qualitative”
  • Paper antibiotic discs are placed on the inoculated Mueller-Hinton agar.
  • Plates are incubated for 16-24 hours and the zones of growth inhibition around each antibiotic disc are measured to the nearest mm
  • The zone of inhibition is measured using a ruler or calliper
A

DISK DIFFUSION or KIRBY BAUER METHOD

47
Q

COMMONLY USED SUSCEPTIBILITY TESTING:

Breakpoints provided by the CLSI standards are
interpreted as?

A

Susceptible, intermediate or resistant

48
Q

COMMONLY USED SUSCEPTIBILITY TESTING:

paaral nalang ng CLSI guidelines thx

49
Q

COMMONLY USED SUSCEPTIBILITY TESTING:

TOF. For vancomycin, there is no recommended breakpoint because
the only acceptable way of susceptible testing for this antimicrobial is through the MIC.

50
Q
  • Plates can be read manually or automatically
  • Automated reader (ID and AST)
  • It takes a picture of each well which can emit fluorescence or turbidity. It can generate an ID and AST using the software
  • Fast and accurate plate reader for maximum consistency
  • Quickly report results
A

Sensititre Optiread.

51
Q
  • Perform manual readings from Sensititre antimicrobial susceptibility testing MIC plates
  • The easy-to-read digital plate image allows users to read and automatically record the MIC result
  • Digital MIC viewing system
A

SENSITITRE VIZION

52
Q

SENSITITRE VIZION

Interpret the results
* Blue
* Red

A

■ Color red (positive) - means growth
■ Color blue (negative) - no growth

53
Q
  • Full walkaway automation
  • When incubating plates, the machine will read its barcode and automatically read the results the following day.
  • Automatically incubate and read plates for identification and susceptibility.
A

SENSITITRE ARIS2X, HIQ

54
Q

TOF. ARIS2X has a higher capacity (60-100) and you can load by batch,
while the HIQ can only be loaded 1 by 1.

A

F (HIQ then ARIS2X)

55
Q

● A single-screen result
● Has an expert system: shows warning, rules, and modifications
● Based on the CLSI, EUCAST, and FDA updates
● Full cascading and suppression

A

SENSITITRE SWIN SOFTWARE

56
Q

SENSITITRE SWIN SOFTWARE

pls study how to interpret it

57
Q

Summarize the process

A

Inoculum
Inoculation
Incubation
Read
Interpret

58
Q

DOCUMENTATION VIDEO

  • Offers a number of semi-automated and automated solutions to enhance and streamline the microbiology laboratory workflow
A

ThermoScientific Sensititre System

59
Q

DOCUMENTATION VIDEO

After creating a 0.5 McFarland suspension, measure its
accuracy using?

A

ThermoScientific Sensititre Nephelometer

60
Q

DOCUMENTATION VIDEO

ThermoScientific Sensititre Nephelometer:

  • If your sample is in the red zone
  • If your sample is within the Green Zone

Interpret

A
  • the bacteria count is either too high or too low
  • your suspension is equivalent to a 0.5 McFarland standard
61
Q

DOCUMENTATION VIDEO

After inoculation, seal, scan and place your plate on what
automated reading and incubation system, or incubate offline?

62
Q

DOCUMENTATION VIDEO

  • offers both semi-automated and fully automated options for quickly and accurately reading your MIC plates
A

Sensititre system

63
Q

DOCUMENTATION VIDEO

  • Enhance manual reading
  • allowing users to visually read and record MIC results with customizable lighting options, touch screen navigation and the powerful SWIN Software system
A

Sensititre vizion

64
Q
  • automated fluorometric plate reading system
  • provides fast, accurate plate reads for maximum consistency
  • For full automation, utilize the ARIS 2x system
  • The ARIS 2x incorporates Optiread fluorescence technology along with automated plate incubation for the ultimate automated solution
A

ThermoScientific Sensititre Optiread

64
Q
  • All of our semi-automated and fully automated plate reading options utilize this
  • can easily consolidate your
    entire test program for robust reporting, with the added benefits of LIS connectivity and a very customizable expert system
A

ThermoScientific Sensititre SWIN Software System.

Seminar 1 (5 decks)

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