1. PCR-based tools for the detection of food-borne pathogens

2. PCR market
PCR technology, revolution in microbiological analysis offering:
Reliable results
Greater efficiency and speed in obtaining the results
Easy methodology
Reproducible results
Lower costs

3. PCR market In Europe, 2006 (R+D and clinical applications only):
PCR reagents market: 624 M $
Thermal cycler market: 423 M $
Source: Frost & Sullivan, 2008
Areas of interest for pathogen detection:
Source: adapted from Biosensors and Bioelectronics, Vol. 22, (2007)

4. PCR market Growing interest in rapid methods based on PCR technology
Source: ISI web of science Adapted from Biosensors and Bioelectronics, Vol. 22, (2007)

5. PCR market
Growing number of publications focused on pathogen detection based on PCR technology
Source : ISI web of science Publications on pathogen detection methods over the last 20 years

6. PCR market
CEN Defines the following criteria for a standard PCR method:
Analytic and diagnostic precision
High detection probability
Robustness
Internal amplification control
Low contamination risk
Easy use and interpretation

7. Industry Problems
Food Industry main problems in Quality Control are caused by
Slowness in obtaining the results which causes a delay in the release of analyzed products
Lack of personnel trained to perform PCR-based analyses
Quality Control costs are high
Due to the high number of hours of personnel required per analysis
Due to long time to market product readiness
Pathogens in food samples are usually hard to detect since they are
Present at low levels, with accompanying bacterial flora
Quiescent due to thermal, radiation, freezing treatments…

8. Industry Needs Food Industry requires Fast results
Reliable and robust results
Sensitive methods with high specificity
Easy-to-perform analytical tools
Access to PCR information and training

9. Solutions
Culture & PCR: both based on exponential amplification:
Culture
PCR 1 2 3 4 2n 2n

10. Microbial
Mission:
Develop and produce bio-technological products to provide better, faster, easier, and cost-effective PCR analysis to industry

11. Microbial Solutions HIGHER SPECIFICITY AND SENSITIVITY
No false negatives nor false positives
Influence of accompanying bacterial flora is lower than in traditional culture or immunological systems
Increased sensitivity with a standard pre-enrichment step

12. Microbial Solutions HIGHER THROUGHPUT
Every reaction includes an Internal Amplification Control
1 reaction = 1 analysis No need of parallel control PCR reactions
Different analysis in the sample plate are possible
96 well plate
94 sample analysis
1 Positive control
1 Non Template Control

13. Microbial Solutions EASY TO PERFORM METHODOLOGY
Minimum number of steps
Several pathogens analyzed in one thermocycler run
SAMPLE
ENRICHMENT DNA EXTRACTION REAL-TIME PCR
Using Microbial DNAready kit or other systems
Automation is possible
A good purification of your DNA will avoid inhibitions!

14. Salmonella + Listeria monocytogenes Bactoplex (DNA extraction + PCR)
Microbial Solutions
LOWER COSTS:
Price per analysis:
Salmonella (ISO 6579:2002) and Listeria monocytogenes (ISO 11290:2004) versus Bactoplex (Salmonella + L. monocytogenes)
Salmonella + Listeria monocytogenes
Bactoplex (DNA extraction + PCR)
Positive result
aprox. 24 €
aprox. 12,5 €
Negative result
aprox. 13 €

15. Salmonella + Listeria monocytogenes Bactoplex (DNA extraction + PCR)
Microbial Solutions
LOWER COSTS:
Hours of a technician required per analysis:
Salmonella (ISO 6579:2002) and Listeria monocytogenes (ISO 11290:2004) versus Bactoplex (Salmonella + L. monocytogenes)
Salmonella + Listeria monocytogenes
Bactoplex (DNA extraction + PCR)
Positive result
~ 80 min
~30 min
Negative result
~ 50 min

16. Microbial Solutions LOWER COSTS: Time to market product readiness:
Culture (ISO)
DNA extraction + PCR
Positive result
5 days
19 hours
Negative result
3 days

17. Microbial Products Soon available: Sample preparation
Positive controls: Reference material for Real time PCR
PCR Quantitation Standard
Soon available:

18. Microbial Products
Results in less than 4 hours after pre-enrichment in BPW
Detection limit tested: 3ufc/l in water samples and 5ufc/25g in food
With an internal amplification control to avoid false negative results
100% sensitivity and specificity
100% efficiency
Validation against European Standard FOOD-PCR method in dairy products, eggs, raw and cured meat, poultry, raw and smoked fish, species, vegetables, fruits, and precooked dishes.

19. Listeria monocytogenes
Microbial Products
Determination of
Listeria monocytogenes
Results in less than 4 hours after pre-enrichment in Fraser
Detection limit tested: 3-4 ufc/reaction
With an internal amplification control to avoid false negative results
100% sensitivity and specificity
100% efficiency
Validation in dairy products, eggs, raw and cured meat, poultry, raw fish, species, and precooked dishes.

20. Microbial Products
Results in less than 4 hours after pre-enrichments in BPW and Fraser
Detection limit tested: 3-6 ufc/25g for Salmonella, and 3-4 ufc/reaction for Listeria monocytogenes
With an internal amplification control to avoid false negative results
100% sensitivity and specificity
100% efficiency
Validation in dairy products, eggs, raw meat, poultry, raw vegetables, raw and smoked fish, and precooked dishes

21. Microbial Products Results in less than 4 hours
Detection limit: 3-4 ufc/reaction
With an internal amplification control to avoid false negative results
100% sensitivity and specificity (detects all 15 serogroups L. pneumophila)
100% efficiency
Validation against ISO in water sample from showers, refrigeration towers, environmental outbreaks, and water supply systems.

22. Microbial Products Results in less than 4 hours
Quantification limit: 1 ufc/reaction
With an internal amplification control to avoid false negative results
100% sensitivity and specificity (detects all Legionella species)
100% efficiency
SYBRGreen based detection, easy interpretation of the results by dissociation curves.

23. Microbial Products Results in less than 4 hours
Detection limit: 1 ufc/reaction
With an internal amplification control to avoid false negative results
100% sensitivity and specificity (detects all thermotolerant Campylobacter)
100% efficiency
Validation against ISO in raw meat, poultry, fruits, vegetables, dairy products, raw fish, and others.

24. Microbial Products Results in less than 4 hours
Detection limit: 1 ufc/reaction
With an internal amplification control to avoid false negative results
100% sensitivity and specificity (detects all Cronobacter species)
100% efficiency
Validation against ISO in powdered infant formula.

25. Microbial Products Results in less than 4 hours
Detection limit: 1 ufc/reaction
With an internal amplification control to avoid false negative results
100% sensitivity and specificity (detects the presence of all Salmonella and Cronobacter species)
100% efficiency
Validation against ISO 6579:2002 and ISO in powdered infant formula

26. Microbial Products Fast DNA extraction Minimum number of steps
Tested on dairy products, raw vegetables, eggs, fruits, fruit juices, precooked dishes, raw and cured meat, poultry, raw and smoked fish, ice- creams

27. Microbial technical product benefits
OPEN KITS: Microbial products work with most of the thermocyclers in the market.
LOWER COSTS: cheaper analysis, less technician’s hours, shorter time to market.
AVOID RISKS: protect your brand with our carefully chosen oligonucleotides using scientific criteria:
Highest stability / lowest variability
Highest inclusivity and specificity
Avoid false negative results
Microbial Kits 27

28. Microbial support to implement PCR
TURN-KEY SOLUTION
We study your needs depending on the type and number of samples analyzed
We propose the best solution to your needs
We come to your lab and set-up your real-time PCR-based system
We perform the first analysis with you in your lab
We provide courses and training on PCR analysis
We offer full support for any technical problem you may find

29. Microbial
Microbial efforts are exclusively directed to research, development, and production of new tools for the detection of pathogenic bacteria by PCR
Years of experience help in identifying the best genetic targets, and in the design of the appropriate molecular tools to detect them
Quality is our main goal
Our analysis systems are based on years of rigorous investigations, and on the application of the characteristics and conditions that a good molecular target must accomplish.
This has lead us to present a product which goes a step forward in the detection of food-borne pathogens.

30. Microbial Official Supporters
CDTi
CIDEM
PCiT de Girona
AGAUR
Universitat de Girona (UdG)
Generalitat de Catalunya (Catalonia Government)
Ministerio de Educación y Ciencia (Spanish Government)
Ministerio de Industria, Turismo y Comercio (Spanish Government) 30

31. Microbial Certifications
All our products have been validated against International Standard Procedures
UNE EN ISO 9001:2008 (ENAC, UKAS)
AFNOR certification (In progress) 31

32. Parc Científic i Tecnològic de Girona 32
Parc Científic i Tecnològic de Girona
Pic de Peguera, 15. Edifici Jaume Casademont, E
E Girona
Spain
Tel
Fax