P I O N E E R I N G D I A G N O S T I C S

New Opportunities in Pathogen Mitigation

Stan Bailey, PhDSr. Director Scientific AffairsbioMereiux Industry

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Marcel Mérieux1897 - Creation of Institut Mérieux

Dr Charles Mérieux1937 - Dr Charles Mérieux took up the reins

Dr Alain Mérieux1963 - Creation of bioMérieux

Alexandre Mérieux

A HISTORIC FAMILY COMMITMENT TO MEDICINE AND PUBLIC HEALTH WORLDWIDE

A pasteurian tradition: Marcel Mérieux worked with Louis Pasteur in 1894.

Marcel Mérieux,student of Louis Pasteur in 1894.

A commitment that transcends generations:Ever since the creation of Institut Mérieux, a generation has worked one after another to expand its legacy.

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OUR FACTS & FIGURES

Sales amounting to

€2.421 billion

A large installed base of

86,900 systems

90% of international sales

About

11,000employees worldwide

1,600 peopleworking at

20 R&D centers

2 LEADERSHIP POSITIONS IN

MICROBIOLOGY GENERATE 2/3 OF SALES

+ 20 YEARS OF EXPERTISE IN INDUSTRIAL MICROBIOLOGY CONTROL

+ 50 YEARSOF EXPERTISE IN

IN VITRO DIAGNOSTICS

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CHANGE IS THE NEW CONSTANT..

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MOORE’S LAW

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TRANSLATION…?

[Today] you would be able to go with that car

300,000 miles per hour. You would get two

million miles per gallon of gas, and all that for

the mere cost of 4 cents!

-Brian Krzanich, Intel CEO

https://www.nytimes.com/2015/05/13/opinion/thomas-friedman-moores-law-turns-50.html

1971

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WHAT DOES NORMAL LOOK LIKE?

FOOD INDUSTRY CUSTOMER CONSUMERSUPPLIER

RAW MATERIALSPRODUCTION

FINISHED PRODUCT

Maintenance

CONSUMER EXPERIENCECONFORMITY

DATA

RAW MATERIAL

VERIFICATIONIN-PRODUCTION DATA

FINISHED PRODUCT TESTING

MAINTENANCE DATA

ENVIRONMENTAL MONITORING DATA

R&DR&D DATA

COMPLAINTS

ENVIRONMENT

ENVIRONMENT MONITORING DATA

Laboratory Testing

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COMPONENTS OF LINEAR TESTING..

Total Solution Partner

Performance Solutions and Lab Optimization

Optimized space layout and sample flow

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CHALLENGES: LINEAR TESTING

• Data in silos with limited scope i.e. lot/situation specific

• Heavy reliance on sampling plans

• “Plateau effect” from technology adding value

• Proactive is still not predictive

• Mundane: often comes down to meeting regulatory compliance

• Answers the What but not “WHY” and definitely not “HOW”

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LABORATORY TRANSFORMATION

Laboratory• Advanced and

comprehensive analysis

• Centers for Predictive Analytics

In field testing:• Real-time data • Actionable • Less reliance on sampling plans• Usually simple workflows• Cost effective • Labs are pushing out the testing

VIDAS

V itek

I mmuno

D iagnostic

A ssay

S ystem

System Overview

VIDAS® IMMUNO ASSAY

CAPTURE OFANTIGENS

SANDWICH TEST DETECTION

The antibody or proteincaptures the target

pathogens

A second antibodyconjugated with an enzymebinds to specific antigens

The intensity of thereaction is interpreted

by the system

370 nm 450 nm

VIDAS evolution

LMO2 LSXNextDay

SET2

Fragment Fab ELFA

1992 1996

ImmunoConcentration

2002 2003 2004 2006

LDUOICS+ SLM

Listeria

2009

SLMXLMX

1993

ECO

2008

ECPT

Protéine Recombinante de phage

2007

VIDASHeat & Go

GENE-UP®

BioMérieux Real Time PCR Solution For Foodborne Pathogen Detection

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GENE-UP ADVANTAGES

3 Levels of Specificity with FRET Technology Internal Control with every test Rapid, Standardized Workflow for all assay parameters

- 30 minutes sample prep time for 96 tests

Ultra Fast and Efficient mechanical lysis procedure Only two enrichment broths used:

- BPW for GENE-UP E. coli O157:H7, STEC, and Salmonella spp- LPT Broth for GENE-UP Listeria spp. and L. monocytogenes

Assay Mix & Match Capability Open Platform

- Capability for foodborne virus detection, GMO testing, allergen testing, etc.

Remote Access Software Room Temperature reagent storage bioMérieux Service and Support

- Scientific Affairs validation/verification services- Applications support

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Amplification & Detection

SPIN DOWN(10 sec)

ENRICHMENT in X mL Broth

Selective Broth

ConfirmationSample Prep

Transfer 5µl Lysate intoPCR TUBES

PCR MIX (Blue Dye)

Transfer20µL into

LYSIS TUBES(Red Dye)

MECHANICAL LYSIS(5 min) PCR

(<1 h)

GENE-UP ROUTINE METHOD: SIMPLIFIED WORKFLOW

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FRET TECHNOLOGY PRINCIPLE

Target

640 nm detection

470 nm light

Internal Amplification

Control

705 nm detection

470 nm light

640 nm detection

705 nm detection

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GENE-UP PROBE-BASED MELT PEAK ANALYSIS

Melt peak analysis is more sensitive than other real time PCR and is

valuable for detecting low level samples.

IncreaseTemperature

Cool PCR reaction

Probes bind DNA

Probes melt off DNA

High Fluorescence

Low Fluorescence

negative derivative

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GENE-UP PROBE-BASED MELT PEAK ANALYSIS

Sample 13 amplified poorly during PCR but was called positive by melt and confirmed by culture

Low level sample + negatives

Amplification Curves

Melting peaks

This is Confidence

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• Amplifies all O157 isolates target DNA (and not from other E. coli)

• Probe melting peak analysis differentiates H7 (SNP) from non-H7 isolates

GENE-UP PERFORMANCE: E. COLI O157:H7 ASSAY

Inc. Temp

Cool PCR Reaction

Probes bind DNA

Probes melt off DNA

High Fluorescence

Low Fluorescence

Inc. Temp

Cool PCR Reaction

O157:H7 O157:non-H7

~ 62 C ~ 58 C

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O157:H7O157: non-H7

E. coli O157:H7 and O157:non H7

isolates can be amplified

GENE-UP PERFORMANCE: E. COLI O157:H7 VS. O157:NON H7

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GENE-UP VALIDATED MATRICES

GENE-UPSalmonella spp.(AOAC)

Fresh Raw ground Beef (25g & 375g)Raw beef trim (25g & 375g)Raw ground pork (375g)Raw ground bison (375g)Fresh Raw chicken breast (25g)Raw ground chicken (25g)Chicken carcass rinse (30mL)Fresh Raw fish (25g)Creamy peanut butter (25g & 375g)Vanilla ice cream (25g)Dry pet food (25g & 375g)Pasteurized liquid whole egg (25g & 100g)Powdered egg (100g)Cooked ham (25g)Lettuce (25g)Chocolate (25g)Whey protein powder (375g)Garlic powder (375g)Onion powder (375g)Dark chocolate (375g)Stainless steel (sponge)

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Work Flow

AOAC RI ValidatedProtocol

FOOD 25g

AOAC RI ValidatedProtocol

FRESH BEEF 25g

AOAC RI ValidatedProtocol

FRESH BEEF 375g

AOAC RI ValidatedProtocol

DRY PET FOOD 375g

GENE-UPSalmonella

GENE-UPSalmonella

GENE-UPSalmonella

GENE-UPSalmonella

Enrichment Broth BPWPre-warmed

BPWPre-warmed

BPW (1:4)Pre-warmed

BPW (1:4)

Enrichment Time42°C for

18-24 h42°C for

8-24 h42°C for

10-24 h42°C for

22-26 h

Sample Prep Time 25 min 25 min 25 min 25 min

No Pipetting Steps 4 4 4 4

Run Time on System 58 min 58 min 58 min 58 min

Min TTR 19.5 h 9.5 h 11.5 h 23.5 h

GENE-UP WORK FLOW & TIME-TO-RESULT

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WHAT DOES NORMAL LOOK LIKE?

Linear testing Dynamic testing

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$1K GENOME WITH $$$ COST ANALYSIS

https://www.researchgate.net/publication/320044302_

Next gen sequencing

$1K genome

Commun ACM. 2016 Aug; 59(8): 72–80.

Moore’s

Kryder’s

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SEQUENCING APPLICATIONS: TODAY

• Identification• Typing and subtyping• Serotyping • AMR• Virulence typing

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EVOLVING SEQUENCING APPLICATIONS

In Greek “meta” means “transcendent”…..

= Meta.. Genomics • WGS • Amplicon (targeted)• .• .• .• .

= Meta.. Transcriptomics

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SOLVING PROBLEMS: DYNAMIC TESTING

Sequence type

Physiological PhysicalChemical Attributes

S. cer. Var Diastaticus Positive

Sequencing ComputationPredictive analytics &

Probability %

Population structure

SourceSpoilage Shelf lifeOthers…

In-field/Lab test

Laboratory Analysis

MACHINE LEARNING

P I O N E E R I N G D I A G N O S T I C S