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Automation in Microbiology Lab

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Anuj Mishra
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80 views27 pages

Automation in Microbiology Lab

Uploaded by

Anuj Mishra
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPT, PDF, TXT or read online on Scribd
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Automation in microbiology lab.

Dr. Baha Eldin Khalid Elamin


Overview
• Background/history
• Plate streakers
• Automated Urine Analysers
• Blood cultures
• Automated Susceptibility Testing
• Automated ID testing
• Malditof
• TLA
• Molecular assays in bacteriology
• Automation and volume
• Automation and commerce
Bacteriology: The Origins
• 1881-Nutrient Gelatin first demonstrated by
Robert Koch
• 1887 agar plates developed
• 1905 - MacConkey agar first used
• 1900s Dyes used to make media selective
• 1919 - Blood Agar first used to study
haemolysis of streptococci
So what's changed??, not much really

• Agar plates are still the main media for the majority
of microbial sub-culturing and the backbone of the
bacteriology lab.
• Disc diffusion is still used in the vast majority of
laboratories – if not all to some degree
• Automated ID/Sens machines (VITEK and
PHOENIX) have improved – however many of the
concepts are still the same, using broth dilution
breakpoint methodology.
• Development in molecular diagnostics.
What about some other techniques?
• Manual streaking of plates has not really
changed since solid agar plates were first used
• Microscopy is mostly unchanged –
• Incubators while probably more reliable are
essentially the same
• Plate reading has not really changed over years
– although you are not supposed to sniff plates
anymore….but we know it happens
Pre-analytical – Plate Streakers
• Select appropriate media
• Loads the samples
• Spreading the inoculum to obtain isolated
single colonies following incubation
• Suppliers: WASP (Copan) Previ-Isola
(BioMerieux) Innova (BD) and Inoqula
(KIESTRA)
• Not all systems include Gram stain preparation
Previ-isola
Innova
WASP

Inoqula
Automated Urine Analysers
• Automated Dip-strip inoculation and
reading
• Cell counts performed automatically
– either by flow cytometry or (more
recently) high resolution optics
taking pictures of cells
• Present now in many labs
Blood Cultures
• Standard in most diagnostic labs, varying
sizes.
• Better detection times
• More advanced media
• Reduced total incubation before calling a
bottle a final negative
• Suppliers: BacT/Alert (BioMerieux) BACTEC
FX (BD)
Alert

lastest BD FX
Biomeriux lastest Bact
The Automation of Susceptibility Testing

• 1940s started playing with putting antimicrobial


agents into agar, both for selective culture and
susceptibility testing.
• Late 1940s: diffusion techniques on filter paper,
6mm disc 1947
• 1966 Kirby/Bauer attempt to standardise with
their disc diffusion technique
• 1975 This technique becomes the basis for the
NCCLS (CLSI) standards.
The Automation of Susceptibility Testing

• 1974 – First automated AST marketed by


Pfizer called the Autobac
• 1977 Abbott introduce the MS 2 System
• 1977 McDonnell Douglas Corporation launch
the AMS System – this was the predecessor to
the first Vitek Machine
• 1977 First standarised microtitre plates with
Antibiotics introduced – leads the way for
Microscan, Sensititre and BBL Spector
Automated ID and Sens
• Automated ID testing available since 1977
• Biochemical substrates miniaturised and read by
colourmetric or fluormetric means
• Available in many bigger labs
• Multiple Antibiotics in different dilutions available
on Cards or panels to ascertain MIC – Expert
Functions
• Suppliers: Microscan Walkaway (Dade Behring)
Vitek2 (BioMerieux) BD Phoenix (BD)
• Cost neutral on ID part but not on susceptibility
testing. What will happen to these systems with
introduction of Maldi-tof?
Microscan
walkaway

BD Phoenix
Vitek2
MALDI-TOF
• Protein based spectral identification of bacteria
• Identifications available in literally minutes – not
hours

• Tiny amount of bacterial growth needed – not


affected by media or incubation conditions

• Minimal cost per test, virtually no consumables


• Suppliers : BD/Bruker, BioMerieux
Vitek MS

Microflex
What is coming?
• What is next in the world of Microbiology
Automation??
• Some companies are in the process of
producing TLA for Microbiology – similar to
those seen in Biochem/Haem.
• Putting specimens on a track – with no human
intervention until plate reading time – and
even then its not like you know it…
Spanner in the works….
• All of this new automated technology assumes
that bacterial culture on agar plates will remain
the cornerstone of microbiological diagnosis
for the forseeable future.
• However molecular assays becoming more
commonplace….even in bacteriology.
• Virology now becoming increasingly
molecular. Viral culture will eventually
disappear from the diagnostic scene.
Some of the current molecular assays
available commercially for bacteriology
• Chlamydia + gono PCR
• Illumigene (LAMP) for C.difficile toxin, Gp B strep
• PCR for Group B strep and MRSA and VRE & ESBL
enzymes
• PCR Bordetella and C.difficile toxin
• PCR and gene probe for mycobacteria
• DNA probes for enteric specimens.
• DNA probes for candida/BV/trichomonas
• 16sRNA identification
What is in the pipeline for molecular
assays in bacteriology?
• Microarrays
Still in research phase.
Still need extraction and
amplification step.
Why is bacteriology not completely
molecular?
• Cost: Cost of Culture even including labour is
inexpensive.

• Culture allows Quantification of bacteria (but


does it matter that much really?)

• Looking for a range of different pathogens.


Current methods aren’t that good at it. PCR
gives up at about 5.
Will the agar plate eventually
become defunct?
• Yes, but possibly not in my working lifetime.
• The companies producing culture automation
are not daft. If they thought that agar plates
would disappear in the next ten years, then
they would not be R&Ding plate streakers,
smart incubators etc.
• Leasing, not buying outright such equipment
would be prudent.
Automation and Volume
• Automated methods most cost-effective
when handling large volumes of samples.
• Favours larger laboratories.
• Interfacing is key
• Most of the cost savings are in labour.
Automation and Commerce
• Two main market players currently in this
field, BD and Biomeurieux.
• As the number of pieces of automated
equipment in a lab increases, there is more and
more incentive for laboratories to “affiliate”
with a company for all their automation
requirements.
• Interesting times ahead….

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