What is the Future of Pharmaceutical Microbiology?
Tony Cundell, Ph. D.
Consulting Microbiologist,
Scarsdale, New York
September 2015 bioMerieux Meeting 1
Future Trends
September 2015 bioMerieux Meeting 2
• In my opinion, what are the future trends in pharmaceutical microbiology?
• Knowing these trends, how can we response to them?
Trends in Pharmaceutical Microbiology Some recent industry trends that I have noticed include:
• Replacement of Growth-based Microbiology Testing Methods with Non-growth Methods.
• More Emphasis of Quality Risk Management and less on Microbiological Testing.
• Integration of the Human Microbiome Findings into Clinical and Pharmaceutical Microbiology.
• Movement of Microbial Testing from the Laboratory to the Factory Floor or the Patient Bedside.
September 2015 bioMerieux Meeting 3
Trends in Pharmaceutical Microbiology
Industry Trends (Continued):
• Further Implementation of Advanced Aseptic Processing and Robotics
• Replacement of Overkill Sterilization Cycles with Bioburden-based Cycles
• Outsourcing of Routine Microbiological Testing to Contract Testing Laboratories
• Re-evaluation of Pyrogenicity and the role of Bacterial Endotoxins Testing
• Digital Disease Detection
September 2015 bioMerieux Meeting 4
Emerging Microbiological Testing Technologies
• Given the time limitations, this presentation will survey the emerging microbiological testing technologies suitable for the pharmaceutical industry.
• In terms of my background I am an internationally-recognized expert in rapid microbial methods who chaired the PDA task force responsible for the 2000 Technical Report Number 33 on the evaluation, validation and implementation of advanced microbiological methods.
• Also I am member of the 20015-2020 USP Microbiology Expert Committee.
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Areas of Discussion
• Rapid microbial methods suitable for air, surface and water monitoring, pharmaceutical ingredient, intermediate and final product testing, and microbial identification will be highlighted and discussed.
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Compendial Guidance
September 2015 bioMerieux Meeting 7
• USP General Notices
• USP <1223> Validation of Alternative Microbiological Test Methods
• Ph. Eur. 5.1.6 Alternative Methods for the Control of Quality.
Other Guidance
• ICH Guidance Q2 (R1) Validation of Analytical Procedures: Text and Methodology, May 1997
• ASTM Standard E2935 Standard Practice for Conducting Equivalence Testing in Laboratory Application, 2013e
• PDA TR#33 (Revised) The Evaluation, Validation and
Implementation of New Microbiological Methods 2014
September 2015 bioMerieux Meeting 8
Relocation of Microbial Testing
• The movement of microbial testing from the laboratory to the factory floor is under way.
• This trend was influenced by the adoption of online water monitoring for conductivity and Total Organic Carbon.
September 2015 bioMerieux Meeting 9
Relocation of Microbial Testing
• The advantages of factory-floor microbial monitoring of utilities, clean rooms and intermediates are that it promotes better in-process control, eliminates taking samples and conveying them to the lab for testing, employs rapid microbial methods, and reduces product failure.
September 2015 bioMerieux Meeting 10
Relocation of Microbial Testing
• Examples of factory-floor testing include LASER-induced fluorescence bioburden monitoring of air, water and in-process intermediates, hand-held cartridge-based endotoxin testing equipment, and in-process sterility testing.
September 2015 bioMerieux Meeting 11
Origins of Traditional Methods
• C’est un grand progrês, Monsieur” Louis Pasteur to Robert Koch (Pictured below), on observing his demonstration of the plate technique, at the Seventh International Medical Congress, London, August 1881.
September 2015 bioMerieux Meeting 12
Replacement of Traditional Microbial Tests
• Growth-based microbiology methods, i.e., traditional plate counts, sterility testing and microbial identification methods are being replacement with non-growth methods, i.e., Scan RDI (solid phase cytometry), D-count (flow cytometry) and Vitek MS (MALDI TOF mass spectrometry) marketed by bioMerieux.
September 2015 bioMerieux Meeting 13
Water Monitoring • Pharmaceutical companies routinely monitor
incoming potable water, and their systems producing and distributing Purified Water, USP and Water For Injection, USP .
• Typically, product is not released to inventory until the water monitoring results are reviewed and found to be satisfactory.
• Although the standard plate count and membrane filtration methods are satisfactory, incubation times of up to 5 days delays the detection of adverse trends and product release.
September 2015 bioMerieux Meeting 14
Candidate Methodologies
Candidate rapid water monitoring systems and their technology include:
• PallChek System (ATP bioluminescence) • Milliflex Rapid Detection System (ATP
bioluminescence) • Scan RDI Microbial Detection System (Solid Phase
LASER Scanning Cytometry)* • RBD 3000 System (Vital Staining Flow Cytometry) • Growth Direct System (Advanced Imaging of
Microcolonies) • IMD-W (LASER-induced fluorescence)
* Marketed by bioMerieux
September 2015 bioMerieux Meeting 15
Microbial Limit Testing
• Many non-sterile pharmaceutical and over-the-counter drug products are subjected to Microbial Limits Testing prior to release
• The USP microbial enumeration and absence of specified micro-organism tests are protracted and delay product release.
• Since many products have little or no bioburden is it possible to screen products for release?
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Candidate Methodologies Candidate rapid microbial enumeration tests and their
technology include:
• Celsis Rapiscreen (ATP bioluminescence)
• PallChek System (ATP bioluminescence)
• Milliflex Rapid Detection System (ATP bioluminescence)
• D-Count System (Flow Cytometry)*
• Growth Direct System (Advanced Imaging)
• Scan RDI (Solid Phase Cytometry)*
• PTS-Micro (Solid Phase Cytometry)
• Sieve-ID MuScan (Solid Phase Cytometry)
* Marketed by bioMerieux
September 2015 bioMerieux Meeting 17
Microbial Limit Testing
Candidate rapid absence of specified microorganism tests and their technology include:
• Vitek MS (MALDI – TOF Mass Spectrometry)*
• BAX Microbial Detection System (PCR DNA amplification based on primers and probes for each specified microorganism)
• Roche LightCycler PCR Detection System (PCR amplification based on primers and fluorescence hybridization probes)
* Marketed by bioMerieux
September 2015 bioMerieux Meeting 18
Microbial Identification
• With the introduction of MALDI TOF mass spectrometric microbial identification there is an opportunity to establish a new paradigm for routine microbial identification.
• With the materials and labor cost for identification being less than U.S $1.00, all bacterial colonies can be identified directly from the primary isolation plates used for facility, water and product testing to continuously evaluate the microbiota associated with your manufacturing plant.
September 2015 bioMerieux Meeting 19
September 2015 bioMerieux Meeting 20
Cost Comparison of Different Bacterial Identification Systems
Identification
Instrumentation
Capital Cost Estimated Cost per
Test
Turnaround Time
Gram Stain Microscope $0.8 10 minutes
API Strips Incubator $5 Overnight
Omnilog $80-100K $10-20 4-6 hours
VITEK Compact 2 $80-100K $8-12 5-6 hours
Vitek MS (MALDI
TOF Mass
Spectrometer)
$180K $1-2 10-20 minutes
ABI MicroSeq
(rRNA base
sequencing)
$150K $80-120 (U.S.
contract lab)
3-6 hours
MALDI TOF Mass
Spectrometer plus
ABI MicroSeq
$330K $35 (U.S. contract
lab)
5 days
September 2015 bioMerieux Meeting 21
Vitek MS Work Flow
Profiling results from different Bacillus strains
-6000
-4000
-2000
0
2000
4000
6000
a.i.
B. globigii
B. licheniformis
B. subtilis
B. thuringiensis
?
Unknown microrganism
Select a colony
Transfer a thin-layer onto a MALDI target plate
Generate MALDI-TOF profile spectrum
MALDI-TOF Pattern Matching
Identified species Bacillus subtilis
Microbial Limit Testing
Candidate rapid absence of specified microorganism tests (Continued):
• DiversiLab (DNA barcoding and analysis for strain typing)*
• RiboPrinter Microbial Characterization System (Based on rRNA gene restriction pattern determinations for strain typing)
• Gen-Trak Bacterial Detection System (DNA Hybridization).
• Bactometer (Impedence to detect growth in selective media)
* Marketed by bioMerieux
September 2015 bioMerieux Meeting 22
Sterility Testing
Candidate rapid sterility tests and their technology include:
• Scan RDI Microbial Detection System (Solid Phase LASER Scanning Cytometry)*
• BacT/ALERT Microbial Detection System (CO2 sensor)*
• BACTEC System (pH sensor)
• Milliflex Rapid (ATP Bioluminescence)
• easyMAG* & ABI 7500 System (RT-PCR) * Marketed by bioMerieux
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Other bioMerieux Innovations
• Previ (Automated Gram staining system)
• AirIdeal 3P Traceability (Active air samplers, with an integrated network-based system including 2P bar-code scanners and remote embedded software)
• BioBall (High-precision QC microorganism inocula)
• LabGuard 3D (24/7 environmental monitoring system recording temperature, static pressure, humidity and CO2 concentration)
• BacT/ALERT Virtuo (automated blood culturing)
• Advencis (Advanced image processing to read agar plates)
September 2015 bioMerieux Meeting 24
RMM Advantages
• In a recent Wall Street analyst briefing the Charles River Laboratories Executive Jim Jordan emphasized Risk Reduction, Time to Result and Cost Saving when discussing their Celsis acquisition.
• These concepts are applicable to bioMerieux’s microbiology product line.
September 2015 bioMerieux Meeting 25
Risk, Time and Cost Continuums
Parameter RMMAdvantagesRiskMitigation ImprovedDetection
OnlineversusSample-basedTestingContinuousversusPeriodicTestingIn-processversusLaboratory-basedTestingReal-timeDecisionMaking
TimetoResult RealTime(Rapid)1-3HoursTurnaroundOvernightTurnaround24to48Hours5to7DaysTurnaroundAtleast14DaysTurnaround(Slow)
Cost LowversusHighCapitalCostNoReagentsversusReagentsSamplingversusOnlineTestingLowversusHighCalibrationCostsLowversusHighMaintenanceCostsLowversusHighAnalystSkillsandTraining
Technology GrowthDependentversusGrowthIndependent
September 2015 bioMerieux Meeting 26
Unmet Stakeholders Needs
• The sterility testing needs for short-lived products such as compounded sterile preparations, cell based therapies and radiopharmaceuticals are not being meet by the current USP/Ph.Eur./JP sterility test what has at least a 14-day incubation period.
• These products are manufactured and infused into patients before the completion of the sterility test.
• Technologies currently marketed by bioMerieux, i.e., ScanRDI and BacT/ALERT System, are available to meet these needs.
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BioMerieux Sterility Testing Options
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TestAttribute USP<71>Sterility
TestScanRDISystem BacT/ALERTDual
TemperatureSystem
Growth-basedmethod
Yes No Yes
Technology Classical SolidPhaseCytometry
CO2sensor
Media Soybean-caseindigestandthioglycollatebroth
Resuscitationstep
Proprietaryaerobicandanaerobicbroth
Samplevolume AsspecifiedinUSP<71>Table2
Variable 10mL
Timetoresults
Atleast14days 1-2hours 5-7days
Verification Subculture Fluorescencemicroscopy
Subculture
ValidationRequirement
Suitabilitytestingonly
EquivalencytoUSP<71>
EquivalencytoUSP<71>
September 2015 bioMerieux Meeting 29 November 14, 2011 Eastern Analytical Symposium,
Somerset, NJ 29
BacT/Alert Microbial Detection
System
September 2015 bioMerieux Meeting 30 November 14, 2011 Eastern Analytical Symposium,
Somerset, NJ 30
BacT/Alert Microbial Detection
System •LED illuminates sensor
• Continuous monitoring: bottles are
read every 10 min.(144 times/day)
• Photodiode collects reflected light
• Signal transmitted to computer
• “Reflectivity Units” plotted over time
Scan RDI System
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ScanRDI System
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Conclusions
• Although pharmaceutical microbiology is a relatively conservative scientific discipline, due recent advances it is a most exciting time to be a microbiologist.
• Lets embrace these advances for the benefit of our companies and users of our pharmaceutical products.
September 2015 bioMerieux Meeting 33
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