virus detection with virseek...one-step real-time rt-pcr is used to detect the rna of the viruses...
TRANSCRIPT
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!