MICROBIOLOGY LAB AUTOMATION Flashcards
TAT
5-7 days
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)
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)
VITEK
Uses______ methodology for organism identification and_____ method for susceptibility testing.
fluorogenic
turbidimetric
Uses fluorogenic methodology for organism identification and turbidimetric method for susceptibility testing.
VITEK
VITEK
• Can provide ID/AST results in as little as ____ hours.
6 to 8
VITEK
identifation
Susceptibility
Fluorogenic
Turbidimetric
2 types of VITEK
Vitek2
Vitek2 compact
MALDI
MALDI(MATRIX ASSISTED LASER DESORPTION/IONIZATION
TOF
TIME OF FLIGHT)
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)
MALDI-TOF mass spectrometry operates in two primary stages.
MALDI
TOF
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
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
Identification of bacteria, fungi, and other microorganisms by comparing their unique protein profiles.
Clinical diagnostics and pathogen detection.
MALDI-TOF
MALDI TOF
identication and characterization of proteins and peptides.
Post-translational modification analysis.
Proteomics
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
• 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
- Identify and characterize biomolecules based on their molecular weight.
- Provide rapid and accurate mass measurement for large macromolecules that may be difficult to analyze using other techniques.
- 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
