detection, identification and quantification of genetically modified organisms … · 2015. 7....

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 nilson.cesar@agricultura.gov.br

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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