Detection, Identification and Quantification of Genetically Modified Organisms (GMOs): Ministry of Agriculture, Livestock and Food Supply Approach

Transcription

1 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

2 Minister of Agriculture, Livestock and Food Supply Executive Secretariat Agriculture and Livestock Health and Inspection Secretariat General Coordination of Laboratory Support (CGAL) LANAGRO/ GO LANAGRO/ MG LANAGRO/ RS LANAGRO/ PE LANAGRO/ PA LANAGRO/ SP

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4 GMO analyses at MAPA 2 official labs LANAGRO GO and MG ISO accredited; Inspection; Audits; Monitoring; Interlaboratory comparison and RM production. 2 authorized labs Official analyses; ISO accreditation.

5 National Agricultural Laboratory LANAGRO-MG Founded in 1983 in Pedro Leopoldo, Minas Gerais State Plant Diagnostics and GMO analyses since accredited methods (ISO 17025) GMO Analyses, Authorized Labs Audits, Interlaboratory Comparison and Reference Material Production.

6 Network building IN 01, Jan. 16th 2007 IN 57, Dec. 11th 2003 ISO Standards: 21569, 21570, and 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)

7 Organização do MAPA: laboratórios OGM - GMO Research CBIO Departments (Inspection) - Labeling - Non authorized events - Coexistence - Exclusion Zones CGAL LANAGRO Authorized Laboratory

8 What about seeds? No specific regulation in place in terms of percentage Consumer Protection (?) CTNBIO Authorized event is not automatically allowed to be planted Registering necessary Seed cultivar registered with the GM event Recently GMBV resistant bean (EMBRAPA) Authorized and not registered Carlavirus resistance pursued

9 Available techniques Strip test Protein based test ELISA Protein based test PCR/ Electrophoresis DNA based test - Qualitative Real time PCR DNA based test Quantitative (Relative) Digital PCR DNA based test Quantitative (Absolute)

10 MAPA GMO Laboratories Current perspective: Real Time PCR Qualitative and quantitative methods Quick and accurate response Low initial investiment and simple execution Many CRM available and validated methods CRM necessity for each assessed event Expensive reagents per sample (mix and probes) Could have better precision (high measurement uncertainty)

11 MAPA GMO Laboratories Operating capacity: SAMPLES (2015) LANAGRO (GO) Animal and Vegetable derived food and feed/ Leaves of different species samples LANAGRO (MG) - Animal and Vegetable derived food and feed 420 samples

12 MAPA GMO Laboratories Future perspective: DIGITAL PCR Direct analysis 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

13 Real time PCR x Digital PCR Real Time PCR: GMO Analysis DCt Std. Curve Ct 14,00 12,00 10,00 8,00 6,00 4,00 2,00 0,00 0,0 0,1 1,0 10,0 % GM

14 Real time PCR x Digital PCR Digital PCR:

15 Real time PCR x Digital PCR Digital PCR:

16 Real time PCR x Digital PCR Digital PCR:

17 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

18 Obrigado Thank you Gracias facebook.com/minagricultura twitter.com/min_agricultura youtube.com/minagriculturabrasil