MICROBIOLOGY LAB AUTOMATION Flashcards by Kyra Catedral

TAT

5-7 days

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Principle

Identify and differentiate bacteria of based on biochemical tests

It provides a standardized, miniaturized version of existing identification techniques, which were complicated to perform and difficult to read.

The plastic strip holds mini-test chambers containing dehydrated media having chemically defined compositions for each test.

They usually detect enzymatic activity, mostly related to fermentation of carbohydrates pr catabolism of proteins or amino acids by the inoculated organisms

ANALYTICAL PROFILE INDEX (API)

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Bacterial suspensions used to rehydrate each of the wells and the strips are incubated.

During incubation metabolism produces color changes that are either spontaneous or revealed by the addition of reagents.

All positive and negative test results are compiled to obtain a profile number, which is then compared with profile numbers in mercial codebook or logbook (or online) for the determination and identification of the bacterial species.

ANALYTICAL PROFILE INDEX (API)

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VITEK

Uses______ methodology for organism identification and_____ method for susceptibility testing.

fluorogenic

turbidimetric

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Uses fluorogenic methodology for organism identification and turbidimetric method for susceptibility testing.

VITEK

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VITEK

• Can provide ID/AST results in as little as ____ hours.

6 to 8

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VITEK

identifation
Susceptibility

Fluorogenic
Turbidimetric

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2 types of VITEK

Vitek2
Vitek2 compact

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MALDI

MALDI(MATRIX ASSISTED LASER DESORPTION/IONIZATION

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TOF

TIME OF FLIGHT)

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It is a powerful analytical technique used in mass spectrometry to identify and analyze biomolecules such as proteins, peptides, lipids, and other macromolecules

MALDI-TOF (MATRIX ASSISTED LASER DESORPTION/IONIZATION-TIME OF FLIGHT)

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MALDI-TOF mass spectrometry operates in two primary stages.

MALDI

TOF

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A sample is mxed with a chemical matrix that absorbs laser energy

A laser pulse irradiates the sample matrix mixture, causing the matrix to vaporize and carry the

In this process, the _______are ionized (usually by gaining or losing a proton) without ignificant fragmentation

analyte molecules

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MALDI TOF

Functions:

:Converts the biomolecules into ions for mass analysis

: Separates ions based on their mass-to-charge ratio (m/z) using the TOF analyzer.

: Measures the ion signals to create a mass spectrum, which provic detailed information about the molecular weights of the components.

Ionization

Mass separation

Detection

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Identification of bacteria, fungi, and other microorganisms by comparing their unique protein profiles.

Clinical diagnostics and pathogen detection.

MALDI-TOF

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MALDI TOF

identication and characterization of proteins and peptides.
Post-translational modification analysis.

Proteomics

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MALDI-TOF

Advantages

Advantages
High sensitivity and specificity.
Minimal sample preparation.
Rapid and high-throughput capability.
Non-destructive analysis.

MALDI-TOF

Limitations

Restricted to analytes that can form ions in the MALDI process.

Requires carefui matrix selection for optimal ionization.

May face challenges with complex mixture or overlapping peaks in spectra

The _______ ________utilizes a colorimetric sensor and reflected light to monitor the presence and production of carbon dioxide (CO2) that is dissolved in the culture medium.

If microorganisms are present in the test sample, carbon dioxide is produced as the organisms metabolize the substrates in the culture medium.

When growth of the microorganisms produces CO2, the color of the gas-permeable sensor installed in the bottom of each culture bottle changes to
yellow.

BACT/ALERT® 3D

Microbial Detection System

The BacT/ALERT Microbial Detection System utilizes a_________ and _________ to monitor the presence and production of _______ that is dissolved in the culture medium.

colorimetric sensor and reflected light

carbon dioxide (CO2)

BacT/ ALERT Microbial Detection System

If microorganisms are present in the test sample,______ is produced as the organisms metabolize the substrates in the culture medium.

When growth of the microorganisms produces CO2, the color of the gas-permeable sensor installed in the bottom of each culture bottle changes to
________.

carbon dioxide

yellow

Used for blood cultures

How many samples???

BacT/ALERT Microbial Detection System

2 (1aerobic; 1anaerobic)

WASP®

• Walk Away Specimen Processor

Is __________

Fully Automated

Automated: Inoculation Streaking Preparation of slides (gram staining) Labelling Disk dispensing

Walk Away Specimen Processor

WASP can…

GRAM SLIDE PREP BROTH INOCULATION DISK DISPENSING TWIN LOOP URINE TURNTABLE

The ___________ is a broth based microdilution method that utilizes a redox indicator (colorimetric oxidation-reduction) to enhance detection of organism growth.

BD Phoenix Automated Microbiology System

• Intended for the in vitro rapid identification (ID) and quantitative determination of antimicrobial susceptibility by minimal inhibitory concentration (MIC) of bacteria.

BD Phoenix Automated Microbiology System

Can be used together with BD Phoenix™ AP for automated inoculum standardization.

BD Phoenix Automated Microbiology System

Pick colonies Prepare McFarland suspension in ID broths Add AST indicator to AST broths and transfer suspension from ID to AST broths Pour the ID and AST broths to each side of the panels Place cap on the panels Scan and place panels in BD Phoenix

BD Phoenix Automated Microbiology System

When microorganisms are present in culture vials, they metabolize nutrients in the culture medium, releasing carbon dioxide into the medium. A dye in the sensor at the bottom of the vial reacts with CO2. This modulates the amount of light that is absorbed by a fluorescent material in the sensor. A photodetector at each station measures the level of fluorescence, which corresponds to the amount of CO2 released by organisms. Then the measurement is interpreted by the system according to pre-programmed positivity parameters.

BD BACTEC

BD BACTEC When microorganisms are present in culture vials, they metabolize nutrients in the culture medium, releasing _____into the medium. A dye in the sensor at the bottom of the vial reacts with CO2. This modulates the amount of light that is absorbed by a fluorescent material in the sensor. A______ at each station measures the level of fluorescence, which corresponds to the amount of CO2 released by organisms. Then the measurement is interpreted by the system according to pre-programmed positivity parameters.

carbon dioxide photodetector

come in gamma-irradiated packs for isolator systems. are for the isolation of microorganisms from samples or specimens. are ideal for cultures requiring prolonged incubation.

BD BBL Isolator Pack plated media BD BBL prepared plated media BD BBL prepared tubed media

• Used for ID/AST • Automatic incubation, test interpretation, and reagent control frees staff for other tasks.

MICROSCAN WALKAWAY

• Principle Uses Colorimetric Optics System consisting of a color wheel/lamp assembly and a Photosensor. There is an initial read at 2.5 hours with a possible final read at 4.5, 5.5, 6.5, 8, 12, 16, or 18 hours (overnight instrument readings, manual readings) depending on the growth rate of the organism being tested.

MICROSCAN WALKAWAY

MICROSCAN WALKAWAY • Principle Uses_________ consisting of a color wheel/lamp assembly and a Photosensor. There is an initial read at____ hours with a possible final read at______ to ______(overnight instrument readings, manual readings) depending on the growth rate of the organism being tested.

Colorimetric Optics System 2.5 4.5, 5.5, 6.5, 8, 12, 16, or 18 hours

1. Identify and characterize biomolecules based on their molecular weight. 2. Provide rapid and accurate mass measurement for large macromolecules that may be difficult to analyze using other techniques. 3. Perform high-throughput analysis for complex samples in various fields, especially microbiology and proteomics.

MALDI-TOF

Applications 1. Microbiology: 1. Identification of bacteria, fungi, and other microorganisms by comparing their unique protein profiles. 2. Clinical diagnostics and pathogen detection. 2. Proteomics: 1. Identification and characterization of proteins and peptides. 2. Post-translational modification analysis.

Vitek MALDI-TOF

• The ______is an advanced tool used for identifying microorganisms and analyzing their metabolic profiles based on their utilization of various carbon sources, nitrogen sources, and other substrates.

Biolog Microbiology Analyzer

Principle • operates on the basis of ***metabolic fingerprinting***

Biolog Microbiology Analyzer

Purpose: • Identify Microorganisms • Characterize Metabolism • Support Research and Development

BIOLOG MICROBIOLOGY ANALYZER