MILLIFLEX RAPID

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MILLIFLEX RAPID

• Rapid detection of microorganisms

Dr. Frank Panofen Millipore Corporation
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Rapid Microbiology Outline

• Generell Considerations
• Milliflex Rapid Product Overview
• Applications

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Rapid Microbiology

• Long response times of traditional methods
• Fast detection
• Reliable Filling
• Raw Materials
• Results in-time
• CIP
• Money saving
• Refusal of Raw Material
• Troubleshooting (contaminated filling-lines)
• Control over Process through just-in-time results

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Rapid Microbiology Implementation I

• Know your current method
• Product sample (filtrability, inhibitory
  properties)
• Preparation conditions (MF, Spread plates)
• Target MO
• Typical counts
• Number of samples
• Incubation conditions (Media, time, temperature)
• Cost

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Rapid Microbiology Implementation II

• Define Requirements
• Time Frame (Sampling Detection)
• Operational Requirements
• Membrane filtration
• Performance (e.g. Viability, Throughput.....)
• Results delivery
• Life cycle of the Method
• Microorganisms (Y M, BAC)
• Cost
• Documentation/Training/Support

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Rapid Microbiology Implementation III

• Evaluate A Product
• Use the Web, Conferences, Journals, Trade
  Magazines, Trade-shows, etc. to find products
• Request a method development/application study be
  performed with your product sample.
• Evaluate a Vendor
• Support (Availability, Knowledge, Training,
  Response time, Product Life Cycle)
• Repair and Maintenance

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Rapid Microbiology Product Offering

• Until 2003 Microstar
• 2004 Milliflex Rapid

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MILLIFLEX RAPID SYSTEM
Filtration
Milliflex plus pump Milliflex funnel RMDS Membrane
Data displaying and saving
User software
Incubation
Pre-filled Milliflex cassette
Detection
Detection tower Image generator Calibration tool
Reagent dispensing
Reagent Autospray Station
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Sample Preparation Filtration

• We use all the benefits of the Milliflex system
  (only filtrable products can be tested)
• Funnel
• Hardware MilliflexPlus Pump
• Prefilled media cassettes

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Rapid Microbiology Incubation

• Bacteria Microcolonies (102 cells) needed
• Y M down to one cell enough for detection

Number of Organisms
Typical Growth curve
106
Overall time for testing reduced to 1/3 1/4
compared to trad. method.
102
Time (days)
T1
T5
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Incubation time An example

• Results E. coli

Mean of 5 replicates

• 6h time to detection for E. coli

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Application of reagents new autospray station

• Design and efficient dispensing system
• Fully automatic Good uniformity of reagents
• One Spraystation for both reagents
• User friendly interface for
• Routine customer use
• cleaning and customer maintenance
• repair and preventive maintenance
• Calibration assistance

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Spray Quality
Efficiency 70

• High efficiency

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Milliflex Rapid Basic Technology overview

• ATP D-luciferin Oxygen (O2)
• AMP PPi Oxyluciferin CO2

Luciferase Mg2
LIGHT

• Wavelength of emitted photons 562 nm
  (green-yellow).
• Number of photons proportional to quantity of ATP
  in the micro-organisms.
• Few thousand photons/s
• High sensitivity technology needed

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ATP Bioluminescence

• The presence of ATP serves as a marker for cell
  viability
• Found in all living cells
• ATP is not associated with dead cells
• ATP can be rapidly released from microbial cells
  and accurately measured
• Cellular ATP Levels Depend Upon
• Type of microorganism Yeast gt bacteria
• Metabolic state of microorganism Healthy gt
  Stressed gt Spore
• ATP Bioluminescence can detect 1cfu/sample

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Milliflex Rapid Technology overview

• Image Intensifier Controller
• Provides the high voltage, during the detection

• Computer w. Milliflex Rapid software
• Collect, treat, register data

• Image processor (IQV50) and CG Board
• Collect the accumulated signal from the CCD
  camera.
• Applies discrimination parameters to subtract
  noise from the signal.
• Generates final digital image which is
  transmitted to the computer.

• Detection Tower / CCD Camera
• collects data

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Milliflex Rapid Technology overview
The Milliflex Rapid detection system collects,
amplifies, and registers on a CCD camera the
light activity of bioluminescent reaction.
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Milliflex Rapid Raw data

• And the resulting image is

• The same in 3 dimensions

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Milliflex Rapid Software Data treatment
counting colonies
Parameters (intensity and size of signal vs
background) set in the system (Tower and
software) allow discrimination between true
signals and background
The detection limit of the system is 1 cfu
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Milliflex-Rapid Incubation time

• Yeast and mold varies from 0 - 4 h depending on
  the microorganism and their metabolic state
• Bacteria 1/3 1/4 of the time needed for the
  traditional MF-Method.

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Milliflex Rapid Calibration

• Light source kit

• Calibration module software

• Certificate

• Certificate

• Stable light source linked to international
  standard
• Portable for on site calibration

• Software driven
• Easy to use

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Milliflex Rapid Services

• Setup and Training
• Validation Guide
• Method development
• On-site validation IQ/OQ support PQ
• Maintenance contracts
• preventive and calibration
• Milliflexplus-pump, Spraystation, Detection Tower

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Evaluated Applications Beverage Experiences

• Feasability studies done on
• Ice Tea
• Filtrable Juices
• Water
• Flavoured Water
• Wine
• Cider
• Beer
• Alcopops

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What to know

• Milliflex rapid
• Results are enumeration (CFU) within a few hours
  (yeast and mold) or 1/3 to 1/4 of the time of
  traditional methods (bacteria), with equivalent
  results (CFU)
• Limit of detection 1 CFU / volume filtered
• Low contaminated, filtrable samples
• Ease of use and throughput which comes from using
  the industry standard Milliflex sample
  preparation system.
• Services for all steps of implementation

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Resume

• Rapid Technologies are ready for implementation
• Understand Actual Method
• Evaluate Requirements
• Evaluate Products (Milliflex Rapid)
• Evaluate Vendor (Millipore)

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Millipore Milliflex Rapid System

• Thank you
• for your attention!!!!