Evaluation of Systems for Nopaline Synthase Terminator in Fast and Standard Real-Time PCR to Screen Genetically Modified Organisms

详细信息    查看全文

• 作者：Elisa Pierboni ; Ludovica Curcio ; Gloria Raquel Tovo…
• 关键词：T ; nos (nos ; terminator) ; Genetically modified organism ; Fast real ; time PCR ; Standard real ; time PCR
• 刊名：Food Analytical Methods
• 出版年：2016
• 出版时间：April 2016
• 年：2016
• 卷：9
• 期：4
• 页码：1009-1019
• 全文大小：444 KB
• 参考文献：Arumuganathan K, Earle ED (1991) Nuclear content of some important plant species. Plant Mol Biol Report 9:208–218CrossRef
  Bahrdt C, Krech AB, Wurz A, Wulff D (2010) Validation of a newly developed hexaplex real-time PCR assay for screening for presence of GMOs in food, feed and seed. Anal Bioanal Chem 396:2103–2112CrossRef
  Barbau-Piednoir E, Lievens A, Mbongolo-Mbella G, Roosens N, Sneyers M, Leunda-Casi A, Van den Bulcke M (2010) SYBR® Green qPCR screening methods for the presence of “35S promoter” and “NOS terminator” elements in food and feed products. Eur Food Res Technol 230(3):383–393. doi:10.​1007/​s00217-009-1170-5 CrossRef
  Broeders S, Huber I, Grohmann L, Berben G, Taverniers I, Mazzara M, Roosens N, Morisset D (2014) Guidelines for validation of qualitative real-time PCR methods. Trends Food Sci Technol 37(2):115–126. doi:10.​1016/​j.​tifs.​2014.​03.​008 CrossRef
  Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, Mueller R, Nolan T, Plaffl MW, Shipley GL, Vandesompele J, Wittwer CT (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55(4):611–622CrossRef
  CAC/GL 74-2010. Codex Committee on Methods of Analysis and Sampling. Guidelines on performance criteria and validation of methods for detection, identification and quantification of specific dna sequences and specific proteins in foods.
  Cheng XY, Jun Z, Ka MC, Xiao KL, Yan H (2008) Quantitative real-time PCR assay to detect transgene copy number in cotton (Gossypium hirsutum). Anal Biochem 375(1):150–152CrossRef
  Dörries HH, Remus I, Grönewald C, Berghof-Jäger K (2010) Development of a qualitative multiplex real-time PCR kit for screening of genetically modified organisms (GMOs). Anal Bioanal Chem 396:2043–2054CrossRef
  ENGL (2015) Definition of minimum performance requirements for analytical methods of GMO testing. 20 April 2015. http://​gmo-crl.​jrc.​ec.​europa.​eu/​guidancedocs.​htm Accessed 30 June 2015
  ENGL (2011a) Overview on the detection, interpretation and reporting on the presence of unauthorised genetically modified materials, EUR 25008 EN ISBN 978-92-79-21800-2 ISSN 1831-9424. doi:10.​2788/​89665 Luxembourg, OPOCE
  ENGL (2011b) Verification of analytical methods for GMO testing when implementing interlaboratory validated methods, EUR 24790 EN ISBN 978-92-72-19925-7 ISSN 1831-9424 doi:10.​2788/​88038 89665 Luxembourg, OPOCE
  EURL-GMFF ENGL (2011) Compendium of reference methods for GMO analysis. EUR 24526 EN http://​gmo-crl.​jrc.​ec.​europa.​eu/​gmomethods/​docs/​GMO-JRC_​Reference_​Report_​2011.​pdf . Accessed 14 Nov 2014
  European Council (2007) Regulation (EC) 834/2007 on organic production and labelling of organic products and repealing Regulation (EEC) No. 2092/91. Official Journal of the European Union L 189, pp 1-23
  European Parliament (2003a) Commission Regulation (EC) 1829/2003 of the European Parliament and of the Council of 22 September 2003 on genetically modified food and feed. Official Journal of the European Union L 268, pp 1-23
  European Parliament (2003b) Commission Regulation (EC) 1830/2003 of the European Parliament and of the Council of 22 September 2003 concerning the traceability and labelling of genetically modified organisms and the traceability of food and feed products produced from genetically modified organisms and amending Directive 2001/18/EC. Official Journal of the European Union L 268, pp 24-28
  European Parliament (2004) Commission Regulation (EC) 882/2004 of the European Parliament and of the Council of 29 April 2004 on official controls performed to ensure the verification of compliance with feed and food law, animal health and animal welfare rules. Official Journal of the European Union L 165, pp 1-141
  European Parliament (2011) Commission Regulation (EU) 619/2011 of 24 June 2011 laying down the methods of sampling and analysis for the official control of feed as regards presence of genetically modified material for which an authorization procedure is pending or the authorization of which has expired. Official Journal of the European Union L 166, pp 9-15
  European Parliament (2015) Directive (EU) 2015/412 of the European Parliament and of the Council of 11 March 2015 amending Directive 2001/18/EC as regards the possibility for the Member States to restrict or prohibit the cultivation of genetically modified organisms (GMOs) in their territory. Official Journal of the European Union L 68, pp 1-8
  Guo J, Yang L, Chen L, Morisset D, Li X, Pan L, Zhang D (2011) MPIC: a high-throughput analytical method for multiple DNA targets. Anal Chem 83:1579–1586CrossRef
  Holst-Jensen A, Ronning SB, Lovseth A, Berdal KG (2003) PCR technology for screening and quantification of genetically modified organisms (GMOs). Anal Bioanal Chem 375:985–993. doi:10.​1007/​s00216-003-1767-7
  Holst-Jensen A (2009) Testing for genetically modified organisms (GMOs): past, present and future perspectives. Biotechnol Adv 27:1071–1082CrossRef
  Holst-Jensen A, Bertheau Y, de Loose M, Grohmann L, Hamels S, Hougs L, Morisset D, Pecoraro S, Pla M, Van den Bulcke M, Wulff D (2012) Detecting un-authorized genetically modified organisms (GMOs) and derived materials. Biotechnol Adv 30(6):1318–1335. doi:10.​1016/​j.​biotechadv.​2012.​01.​024 CrossRef
  Huber I, Block A, Sebah D, Debode F, Morisset D, Grohmann L, Berben G, Stebih D, Milavec D, Zel J, Busch U (2013) Development and validation of duplex, triplex, and pentaplex real-time pcr screening assay for the detection of genetically modified organisms in food and feed. J Agric Food Chem 61:10293–10301CrossRef
  ISO 21570 (2013) Foodstuff-Methods of analysis for detection of genetically modified organisms and derived products-Quantitative nucleic acid based methods
  ISO 21571 (2013) Methods of analysis for the detection of genetically modified organisms and derived products-nucleic acid extraction
  ISO 24276 (2013) Foodstuff-Methods of analysis for the detection of genetically modified organisms and derived products. General requirements and definitions
  ISO/IEC 17025 (2005) General requirements for the competence of testing and calibration laboratories
  Madeo L., Pierboni E., Curcio L., Tovo G., Gabrielli F., Rondini C. (2012) . “OGM Sviluppo di una metodica rapida per la tracciabilità della patata su matrici agroalimentari semplici e complesse.” XIV Congresso Nazionale S.I.Di.L.V. Sorrento (Na). 364-366
  Permingeat HR, Reggiardo MI, Vallejos RH (2002) Detection and quantification of transgene in grains by multiplex and real-time PCR. J Agric Food Chem 50:4431–4436CrossRef
  Querci M, Van den Bulcke M, Zel J, Van den Eede G, Broll H (2010) New approaches in GMO detection. Anal Bioanal Chem 396:1991–2002. doi:10.​1007/​s00216-009-3237-3 CrossRef
  Reiting R, Broll H, Waiblinger HU, Grohmann L (2007) Collaborative study of a T-nos real-time PCR method for screening of genetically modified organisms in food products. J Verbr Lebensm 2:116–121. doi:10.​1007/​s00003-007-0189-4 CrossRef
  Ruttink T, Demeyer R, Gulck E, Droogenbroeck B, Querci M, Taverniers I, Loose M (2010) Molecular toolbox for the identification of unknown genetically modified organisms. Anal Bioanal Chem 396:2073–2089. doi:10.​1007/​s00216-009-3287-6 CrossRef
  Stein AJ, Rodriguez-Cerezo E (2009) The global pipeline of new GM crops-implications of asynchronous approval for international trade, EUR 23846 EN ISBN 978-92-79-12603-1 ISSN 1018-5593 doi:10.​2791/​12087 Luxembourg, OPOCE
  USDA (2006) Verification of the Bayer CropScience 35S-BAR method for the detection of LL62/LL601 rice using Real-time PCR, produced by the US Department of Agriculture. http://​gmo-crl.​jrc.​ec.​europa.​eu/​doc/​35SBarRiceVerifi​cationReportrev.​pdf . Accessed 14 Nov 2014
  USDA (2008). United states Department of Agriculture controls over importation of transgenic plants and animals. U.S. Department of Agriculture, O.o.I.G.S.R. Audit report pp 1-26 http://​www.​usda.​gov/​oig/​webdocs/​50601-17-TE.​pdf . Accessed 14 November 2014
  Vaitilingom M, Pijnenburg H, Gendre F, Brignon P (1999) real-time quantitative pcr detection of genetically modified maximizer maize and roundup ready soybean in some representative foods. J Agric Food Chem 47(12):5261–5266CrossRef
  Waiblinger HU, Ernst B, Anderson A, Pietsch K (2008) Validation and collaborative study of a P35S and T-nos duplex real-time PCR screening method to detect genetically modified organisms in food products. Eur Food Res Technol 226:1221–1228. doi:10.​1007/​s00217-007-0748-z CrossRef
  Waiblinger HU, Grohamann L, Mankertz J, Engelbert D, Pietsch K (2009) A practical approach to screen for authorised and unauthorized genetically modified plants. Anal Bioanal Chem 396(6):2065–2072. doi:10.​1007/​s00216-009-3173-2 CrossRef
  Waiblinger HU, Grohmann L (2014) Guidelines for validation of DNA extraction methods applied in subsequent PCR analysis of food and feed products for the presence of genetically modified material. J Verbr Lebensm. doi:10.​1007/​s00003-014-0862-3
  Weighardt F, Barbati C, Paoletti C, Querci M, Kay S, De Beuckeleer M, Van den Eede G (2004) Real-time polymerase chain reaction-based approach for quantification of the pat Gene in the T25 Zea mays event. J AOAC Int 87(6):1342–1355
  
• 作者单位：Elisa Pierboni (1)
  Ludovica Curcio (1) (2)
  Gloria Raquel Tovo (1)
  Martina Torricelli (1) (3)
  Cristina Rondini (1)
  
  1. Laboratorio OGM ed Igiene dell’Ambiente, Istituto Zooprofilattico Sperimentale dell’Umbria e delle Marche, via G. Salvemini n. 1, 06126, Perugia, PG, Italy
  2. Dipartimento di Scienze Agrarie, Alimentari e Ambientali, Università degli Studi di Perugia, Borgo XX giugno 74, 06121, Perugia, PG, Italy
  3. Dipartimento di Medicina Veterinaria, Università degli Studi di Perugia, via San Costanzo 4, 06126, Perugia, PG, Italy
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Food Science
  Chemistry
  Microbiology
  Analytical Chemistry
  
• 出版者：Springer New York
• ISSN：1936-976X

文摘

The increasing number and diversity of genetically modified organisms (GMOs) developed and commercialised forces laboratories to apply many different methods of analysis. Matrix-based approach helps to minimize analytical effort reducing the number of identifications. However, the correctness of screening phase needs efficient methods. In this paper, 15 systems for the nopaline synthase terminator (T-nos) from Agrobacterium tumefaciens, a genetic element present in several genetically modified (GM) plants, were tested. The systems were obtained from three methods, and their primers and probes were combined and tested in real-time polymerase chain reaction (PCR) in fast and standard mode. The results have showed that the fast mode presented the lowest mean quantification cycle (Cq) and the highest ∆Rn, that is, the difference between normalized reporter and baseline. These parameters of system efficiency prove that the fast real-time PCR is a possible approach to obtain good data in less than half the time compared to standard mode. The proposed approach allows a practical way to evaluate the most efficient set of oligonucleotides (e.g., primers and probe) in fast and standard real-time PCR before validation. This article describes also in-house validation of the best set oligonucleotides.