detection, identification and quantification of genetically modified organisms … · 2015. 7....
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Detection, Identification and Quantification of Genetically Modified Organisms (GMOs): Ministry of Agriculture, Livestock and Food Supply Approach
Nilson César Castanheira Guimarães Technical Coordinator – LANAGRO-MG [email protected]
Executive Secretariat
Agriculture and Livestock Health and Inspection Secretariat
Minister of Agriculture, Livestock and Food Supply
General Coordination of Laboratory Support (CGAL)
LANAGRO/GO
LANAGRO/SP
LANAGRO/PA
LANAGRO/PE
LANAGRO/RS
LANAGRO/MG
GMO analyses at MAPA
2 official labs LANAGRO GO and MG – ISO 17025 accredited; • Inspection; • Audits; • Monitoring; • Interlaboratory comparison and RM production. 2 authorized labs • Official analyses; • ISO 17025 accreditation.
Plant Diagnostics and GMO analyses since 2007 – 4 accredited methods (ISO 17025)
Founded in 1983 in Pedro Leopoldo, Minas Gerais State
National Agricultural Laboratory LANAGRO-MG
GMO Analyses, Authorized Labs Audits, Interlaboratory Comparison and Reference Material Production.
Network building IN 01, Jan. 16th 2007 Ü IN 57, Dec. 11th 2003
ISO Standards: 21569, 21570, 21571 and 21572 – DNA Extraction, DNA and Protein Based Qualitative and Quantitaive Analyses;
Minimum Performance Requirements, EURL/JRC and ENGL
Measurement Uncertainty – In-house Development of a Methodology
Interlaboratory Comparison – PRIMAR/LANAGRO-MG – Ground Corn (6 events)
CBIO Departments (Inspection)
CGAL
Organização do MAPA: laboratórios OGM
LANAGRO Authorized Laboratory
- Labeling
- Non authorized events
- Coexistence
- Exclusion Zones
- GMO Research
What about seeds?
Recently – GMBV resistant bean (EMBRAPA) – Authorized and not registered – Carlavirus resistance pursued
Registering necessary – Seed cultivar registered with the GM event
CTNBIO Authorized event is not automatically allowed to be planted
No specific regulation in place in terms of percentage – Consumer Protection (?)
Available techniques
PCR/ Electrophoresis – DNA based test - Qualitative
ELISA – Protein based test
Strip test – Protein based test
Real time PCR – DNA based test – Quantitative (Relative)
Digital PCR – DNA based test – Quantitative (Absolute)
MAPA GMO Laboratories
Quick and accurate response Qualitative and quantitative methods
Real Time PCR
Current perspective:
Many CRM available and validated methods
Low initial investiment and simple execution
CRM necessity for each assessed event Expensive reagents per sample (mix and probes)
Could have better precision (high measurement uncertainty)
MAPA GMO Laboratories
Operating capacity:
SAMPLES (2015) LANAGRO (GO) – Animal and Vegetable derived food and feed/ Leaves of different species 1.685 samples LANAGRO (MG) - Animal and Vegetable derived food and feed 420 samples
MAPA GMO Laboratories Future perspective:
•DIGITAL PCR
• Direct analysis – 20.000 simultaneous reactions
• Nanodroplets/ wells counting (6 a 20 nL) – Flow or Microchip
• Increased precision
• High Initial investiment
• CRMs are unnecessary for quantification (RM certification and versatility)
• Relative low cost of reagents per sample
• Quick and handy
• Vertiginous and continuous evolution of this technique
Real time PCR x Digital PCR Real Time PCR:
GMO Analysis DCt Std. Curve
0,00
2,00
4,00
6,00
8,00
10,00
12,00
14,00
0,0 0,1 1,0 10,0
% GM
∆Ct
Real time PCR x Digital PCR Digital PCR:
Real time PCR x Digital PCR Digital PCR:
Real time PCR x Digital PCR Digital PCR:
MAPA GMO Laboratories Future perspective:
• Increase in workforce – Public contest in 2014
• Enough Equipments and Area
• Increased Operating Capacity and broadened scope Total satisfaction of our clients demands
• RM and CRM production – Official and Authorized Labs
• Continuous Offer of Interlaboratory Comparisons – Official and Authorized Labs and International partners
• International Cooperation – JRC (EURL, IRMM) and RLAC
Obrigado Thank you Gracias
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