Diagnostic Micro Lecture 24

What are advantages of TLA technology in clinical microbiology?

-Closed incubation system -Image analysis and plate inoculation (can be standardized of culture to be more consistent with reporting out results) -Automation reduces processing times especially of negative cultures -Reduces the little things that take up time to better manage work load

What are disadvantages of TLA technology in clinical microbiology?

-Expensive investment (only worth it with high load labs) -Compatibility (WASP and BD-Kiestra aren't compatible with each other; like PC (wasp) to Apple (bd-kiestra)) -Can only handle uncomplicated specimen types

What does the Specimen Processor section of WASP cover?

-Has a HEPA Filtration system so it's safe to culture respiratory specimens - standardized sample created - Whatever tests are ordered for the sample, the system sets everything up(vortex, susceptibility discs, plating on certain media, gram stain (can do it automated or manually), etc.)

What are some advantages of the Phoenix Automated Susceptibility Testing machine?

-Has different panels for GP and GN -Do susceptibilities based on growth -Identification based on many biochemical reactions of carbohydrate and amino acid metabolism -Can be used for susceptibility, ID or both

What are disadvantages of MALDI-TOF tech in clinical microbiology?

-High upfront cost -Not suitable for direct specimens (only good for screening of blood samples; need to follow up with routine culture) -Should not be used for bacteria such as Brucella and Francisella -ID is only as good as the database (limited to library, and how the bacteria proteins are identical to the ones in the database) -E. coli and Shigella sp. cannot be differentiated :(

What are advantages of MALDI-TOF tech in clinical microbiology?

-Unbiased genus and species identification of bacteria, yeasts, and multicellular fungi due to a reliable and large clinical database -Interfaces for Laboratory Information System (LIS) integration -Paradigm shift for diagnostic microbiology -Inexpensive (after initial investment, only costs a couple dollars per organism) -FAST (takes total time of hours/days to identify organism to minutes)

What are some TLA devices?

-WASP (Walk Away Specimen Processor) -WASPLab (whenever a WASP system(s) is connected to other units via conveyer which makes it a TLA) -BD-Kiestra (does same stuff as WASP but is more expensive)

What is the TLA workflow?

-system to plant samples -incubate separately -photograph media to upload to digital system -much more efficient

What is the major benefit of using BioFire - FilmArrays?

A lot quicker identification -Blood Culture Identification panel -Meningitis/Encephalitis panel -Gastrointestinal panel -Pneumonia panel -Respiratory panel (general & a new SARS-CoV-2 addition) Certain panels can ID drug resistant bugs Using multiple steps of PCR, amplification discerning which is which

What are the kinds of organisms don't work as well within Automated Susceptibility Testing?

GPR Streps Anaerobes Non-enterics Staphs & Enterococci

What kind of media is needed for Automated Susceptibility Testing - Phoenix?

Broth diluting testing in order to achieve susceptibilities

What is 16 S Sequencing used for?

From culture, "fast identification" for slow growing organisms

What process isn't eliminated in the workflow by automation the lab?

Isolation of mixed cultures, subjectively pick apart what we think is the predominant bacteria that is causing an infection in our patient

What entails classic clinical bacteriology workflow?

Manually isolating organisms, biochemical arrays, susceptibilities, all with a day in between dependent on if bugs behave/grow in that time period

What does MALDI-TOF MS stand for?

Matrix-Assisted Laser Desorption Ionization - Time Of Flight Mass Spectrometry

What are the procedural steps MALDI-TOF?

Once you have an isolate → spot isolate onto target metal plate (typically with a toothpick or the like) → allow isolate to dry (sometimes treated with formic acid) → following drying, addition of a matrix to then dry as well → put into MALDI and let the lasers do their thing.

What about the WASPLab incubator makes it more efficient than standard swing door incubators?

The WASPLab incubator is a closed system so insertion and extraction of plates are kept at a regulated temperature without exposure to RT (still a maintenance door but want to keep it closed as much as possible)

Why is classic workflow not ideal?

Too slow, not very sensative, labor intensive/requires highly trained individuals to correctly read/perform the tests leading to higher costs

What does TLA stand for?

Total Laboratory Automation