Virus detection with VIR Seek

May 2019

Aaron HuckabeeSales Director North America

Eurofins Technologies

2Eurofins Technologies Marketing_Monthly call

EUROFINS TECHNOLOGIES

Eurofins Technologies history

3Eurofins Technologies

• Launched in 2015 as an International Business Line within the Eurofins Group

• Rapid creation of a network of companies tobuild global expertise and expand product portfolio in the diagnostic test kit market

Key businesses:

• Industrial Solutions: Food, Feed and Environmental Safety

• Animal Health Solutions

• Clinicial Diagnostics

Market Product lines

• Allergens• Food Pathogens • GMOs• Animal Species• Plant Species• Reference Materials

• Algal Toxins• Pesticides• Magnetic Particles• Mycotoxins • Veterinary Drug

Residues

• Viruses• Bacterial pathologies• Farm and companion animals

Method Instruments

• ELISA/CLIA automation

• Infectious Diseases • Autoimmune Disorders• Allergies

Comprehensive product portfolio

Companies

5Eurofins Technologies

Diagnostic kits from farm to fork

Planting Plant Health Animal Health Raw Materials Ingredients & Production & FinishedFeed Processing Products

• GMOs • Plantpathogens

• Environment& Water

• Animal Health• VDRs

• GMOs• Mycotoxins

• Allergens• Pathogens• Animal

Species

• GMOs• Plant

Species

• Allergens• Pathogens• GMOs• Animal

Species

France

BelgiumGermany

Hungary

ItalyUSA

Singapore

Australia / New Zealand

Eurofins Technologies global footprint

6Eurofins Technologies

Spain

Dynamically-expanding global network

India

USADavis Gold Standard DiagnosticsWarminster AbraxisMinnesota ET North America

USADavis Gold Standard DiagnosticsWarminster AbraxisMinnesota ET North America

EUROPEFreiburg GeneScanKassel ImmunolabParis ET FranceMadrid IngenasaTrieste TecnaBudapest ET Hungary

EUROPEFreiburg GeneScanKassel ImmunolabParis ET FranceMadrid IngenasaTrieste TecnaBudapest ET Hungary

ASIAHyderabad AmarSingapore ET SingaporeMelbourne ET Australia

ASIAHyderabad AmarSingapore ET SingaporeMelbourne ET Australia

7Eurofins Technologies

VIRSeek: Virus detection in food

VIRSeek - Background

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Common viruses in food: norovirus, hepatitis A viru s (HAV), hepatitis E virus (HEV), rotavirus

Found on/in food products and in water

Surveillance for foodborne disease outbreaks, Unite d States Annual report (2016)

• Norovirus most common source of single-etiology outbreaks (36 %)

• 17 % of hospitalisations due to HAV

EU summary report on zoonoses, zoonotic agents and food-borne outbreaks (2016)

• 9.8 % of food outbreaks viral

• Caliciviruses caused highest number of illnesses per outbreak

• Norovirus: 31.6 cases/outbreak → 24 % of all illnesses in food outbreaks

VIRSeek - Viruses

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Viral nucleic acid Capsid Nucleocapsid Enveloped Virus

viral nucleic acid capsid nucleocapsid enveloped virus

dsDNA, ssDNSdsRNA, +ssRNA, -ssRNA

circular, fragmented …

empty virus-like particles capsid containing viral nucleic acid

capsid surrounded by an envelope made up of

phospholipids and proteins

infectious non-infectious infectious infectious

Norovirus, Hepatitis A Influenza, HIV

VIRSeek – Our solution

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Challenges for food analysis Low virus concentration in foods

No cultivation possible → concentration steps required

Certain relevant matrices may contain PCR inhibitor s

Viral process controls essential

VIRSeek Solution VIRSeek RNAExtractor Food

VIRSeek Food Norovirus Genogroup I (NoVGI) Real-time RT-PCR

VIRSeek Food Norovirus Genogroup II (NoVGII) Real-time RT-PC R

VIRSeek Food Hepatitis A Virus (HAV) Real-time RT-PCR

VIRSeek Food Murine Norovirus (MNV) / Process Control Virus Real-time RT-PCR

VIRSeek – Solution workflow

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Preparation and concentration of samples according to ISO 15216

VIRSeek RNAExtractor Food

Viral RNA detection (real-time RT-PCR) using :

VIRSeek Food Norovirus Genogroup I (NoVGI) Real-time RT-PCRVIRSeek Food Norovirus Genogroup II (NoVGII) Real-time RT-PCR

VIRSeek Food Hepatitis A Virus (HAV) Real-time RT-PCR

Process control using:

VIRSeek Food Murine Norovirus (MNV) / Process Control Virus Real-time RT-PCR

VIRSeek – Solution components

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VIRSeek RNAExtractor Food

Preparation for Real-time RT-PCR

RNA extraction from viral samples prepared and conc entrated according to ISO 15216

Silica-coated magnetic bead based extraction

Removal of potential inhibitors

Sample types include:• Soft fruits

• Leaf, stem and bulb vegetables

• Bottled water

• Bivalve molluscs

VIRSeek – Solution components

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VIRSeek Food Murine Norovirus (MNV)

Murine norovirus • found in mice and other small animals

• not pathogenic to humans

Process control virus to monitor efficiency of viru s concentration steps

Samples spiked during virus concentration (ISO 1521 6)

Morphologically and physio-chemically similar to pa thogenic viruses• Comparable extraction efficiency

Tested in parallel to each target

VIRSeek – Solution components

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VIRSeek Food Norovirus & HAV kits Norovirus genogroups I & II – most relevant to human health

RNA viruses without an envelope

One-step Real-time RT-PCR is used to detect the RNA of the viruses

IPC is included in order to confirm the validity of each sample

Validated for 3 cyclers:• Agilent AriaMx TM

• BioRad CFX96 Touch TM

• BioRad CFX96 Touch TM Deep Well

• Also validated and certified for kits in the BAC Gene product line

15Eurofins Technologies

THANK YOU!