Bisküvi örneklerinden GDO tespiti için DNA izolasyon metotlarının karşılaştırılması

Öz

Genetiği değiştirilmiş organizmaların (GDO) kullanıldığı yem ve gıda ürünlerinin dünya çapında artış göstermesi, biyoteknolojik yollarla elde edilen bu gıdalar ile, GDO içermeyen gıdalar arasında ayrım yapabilen tespit yöntemlerinin geliştirilmesine yol açmıştır. Farklı ülkelerdeki çeşitli düzenlemelere rağmen, GDO'lu ürünlerin etiketlenmesi konusunda uluslararası düzeyde tutarlı bir yaklaşıma ihtiyaç vardır. Bu sebeple tüketicilere güvenilir ve şeffaf bir gıda güvenliği sağlamak açısından etkili GDO tespit yöntemlerinin geliştirilmesine dair ihtiyaç oluşmaktadır. GDO'ya yönelik doğru ve güvenilir tespit yöntemlerinin oluşturulmasında ilk deneysel işlem basamağı etkili bir DNA izolasyonudur. Bu konuda farklı işlenmişlik seviyesindeki gıdalara özgü DNA izolasyon yöntemlerinin belirlenmesi oldukça önemlidir. Bu çalışmada, bisküvi örnekleri için farklı DNA ekstraksiyon yöntemlerinin karşılaştırılması amaçlanmıştır. Bu sebeple farklı bisküvi örneklerine ait DNA’lar, seçilen üç faklı protokol (CTAB-PVP, modifiye CTAB-1 ve modifiye CTAB-2) kullanılarak izole edilmiştir. İşlenmiş gıda ürünleri olan bisküvilerden ekstrakte edilen DNA örnekleri, DNA konsantrasyonu ve saflığı bakımından değerlendirilmiştir. Elde edilen sonuçlara göre bisküvi örnekleri için hem DNA miktarı hem de saflık oranları bakımından genel olarak en iyi performansı CTAB-2 prosedürü göstermiştir.

Anahtar Kelimeler

• Bisküvi
• CTAB
• DNA ekstraksiyonu
• GDO
• gıda güvenliği

Comparison of DNA isolation methods for GMO detection from biscuit samples

Öz

The global increase in genetically modified organism (GMO) content in feed and food products has necessitated the development of precise detection methods to differentiate between biotechnologically derived foods and those without GMOs. Despite the various regulations in different countries, an internationally consistent approach to labeling GMO products is needed. For this reason, there is a widespread need to develop effective GMO detection methods to provide reliable and transparent food safety to consumers. The first experimental step in creating accurate and reliable detection methods for GMOs is effective DNA isolation. Determining DNA isolation methods specific to different processing levels of foods is very important. This study was aimed to compare different DNA extraction methods in biscuit samples. For this reason, DNA from different biscuit samples was isolated using three different protocols (CTAB-PVP, modified CTAB-1, and modified CTAB-2). DNA samples extracted from biscuits, were evaluated regarding DNA concentration and purity. According to the results obtained, the CTAB-2 procedure generally showed the best performance in terms of both DNA amount and purity rates for biscuit samples.

Anahtar Kelimeler

• biscuit
• CTAB
• DNA extraction
• GMO
• food safety

Kaynakça

1. Abdel-Latif A, Osman G (2017). Comparison of three genomic DNA extraction methods to obtain high DNA quality from maize. Plant Methods 13: 1-9.
2. Aboul-Maaty NAF, Oraby HAS (2019). Extraction of high-quality genomic DNA from different plant orders applying a modified CTAB-based method. Bulletin of the National Research Centre 43(1): 1-10.
3. Abrams SA, Albin JL, Landrigan PJ, Corkins MR, Blanco CL, Fuchs GJ, Spire P (2024). Use of Genetically Modified Organism (GMO)-Containing Food Products in Children. Pediatrics 153(1).
4. Aksoy BT, Ateş Sönmezoğlu Ö (2022). Comparison of modified DNA isolation methods for the detection of GMO in processed foods. International Journal of Life Sciences and Biotechnology 5(3): 546-561.
5. Al-Salameen F, Kumar V, Al-Aqeel, Al-Hashash H, Hejji AB (2012). Detection of genetically modified DNA in fresh and processed foods sold in Kuwait. GM crops & food 3(4): 283-288.
6. Arvas YE, Kocaçalışkan İ (2020). Genetiği değiştirilmiş bitkilerin biyogüvenlik riskleri. Türk Doğa ve Fen dergisi 9(2): 201-210.
7. Arvas YE, Yılmaz K (2019). Genetiği değiştirilmiş bitkilerin biyolojik çeşitliliğe potansiyel etkileri. Yuzuncu Yıl University Journal of Agricultural Sciences 29(1): 168-177.
8. Ashrafi-Dehkordi E, Mazloomi SM, Hemmati F (2021). A comparison of DNA extraction methods and PCR-based detection of GMO in textured soy protein. Journal of Consumer Protection and Food Safety 16(1): 51-57.

9. Arun ÖÖ, Muratoğlu K, Eker FY (2016). The effect of heat processing on pcr detection of genetically modified soy in bakery products. Food and Health 2(3): 130-139.
10. Arun ÖÖ, Yılmaz F, Muratoğlu K (2013). PCR detection of genetically modified maize and soy in mildly and highly processed foods. Food Control 32(2): 525-531.
11. Ateş Sönmezoğlu Ö, Terzi B (2018). Characterization of some bread wheat genotypes using molecular markers for drought tolerance. Physiology and Molecular Biology of Plants 24(1): 159-166.
12. Bøhn T, Millstone E (2019). The introduction of thousands of tonnes of glyphosate in the food chain—an evaluation of glyphosate tolerant soybeans. Foods 8(12): 669.
13. Bitir B, Erkan I, Yükseloğlu EH (2020). Searching of the genetically modified organisms and their products’ status and evaluation of food safety and regulations in Turkey in terms of the Forensic Sciences. Commagene Journal of Biology 4(2): 104-109.
14. Costa J, Melo VS, Santos CG, Oliveira MBP, Mafra I (2015). Tracing tree nut allergens in chocolate: A comparison of DNA extraction protocols. Food Chemistry 187: 469-476.
15. Davison J (2010). GM plants: Science, politics and EC regulations. Plant Science 178(2): 94- 98.
16. Di Pinto A, Forte V, Guastadisegni MC, Martino C, Schena FP, Tantillo G (2007). A comparison of DNA extraction methods for food analysis. Food Control 18(1): 76-80.
17. Elsanhoty RM, Ramadan MF, Jany KD (2011). DNA extraction methods for detecting genetically modified foods: A comparative study. Food chemistry 126(4): 1883-1889.
18. Ergin N, Aydemir SK (2018). Soya bitkisinin hayvan beslenmesindeki yeri ve önemi. International Journal of Eastern Mediterranean Agricultural Research 1(1): 143-157.
19. Erkan I, Dastan K (2017). Real-time PCR detection of genetically modified organisms in several food products and their environmental effects in Turkey. Fresenius Environmental Bulletin 26(4): 2589-2595.
20. Gbashi S, Adebo O, Adebiyi JA, Targuma S, Tebele S, Areo OM, Olopade B, Odukoya JO, Njobeh P (2021). Food safety, food security and genetically modified organisms in Africa: a current perspective. Biotechnology and Genetic Engineering Reviews 37(1): 30-63.
21. Gryson N, Messens K, Dewettinck K (2004). Evaluation and optimisation of five different extraction methods for soy DNA in chocolate and biscuits. Extraction of DNA as a first step in GMO analysis. Journal of the Science of Food and Agriculture 84(11): 1357-1363.
22. Gryson N, Ronsse F, Messens K, De Loose M, Verleyen T, Dewettinck K (2002). Detection of DNA during the refining of soybean oil. Journal of the American Oil Chemists' Society 79: 171-174.
23. ISAAA, (2019). ISAAA Brief 55-2019: Executive Summary Biotech Crops Drive Socio-Economic Development and Sustainable Environment in the New Frontier. 2019. In: https://www.isaaa.org/resources/publications/briefs/55/executivesummary/default.asp [accessed 26.12.2023].
24. Krinitsina AA, Sizova TV, Zaika MA, Speranskaya AS, Sukhorukov AP (2015). A rapid and cost-effective method for DNA extraction from archival herbarium specimens. Biochemistry (Moscow) 80: 1478-1484.
25. Leão-Buchir J, Pedro OIC, Milisse AEM, Mabuie AIM, De-Araujo L, das Neves LI, Correia DA (2022). Detection of genetically modified organisms in food products commercialized in Mozambique. African Journal of Food Science 16(2): 22-29.
26. Li X, Pan L, Li J, Zhang Q, Zhang S, Lv R, Yang L (2011). Establishment and application of event-specific polymerase chain reaction methods for two genetically modified soybean events, A2704-12 and A5547-127. Journal of agricultural and food chemistry 59(24): 13188-13194.
27. Lin CH, Pan TM (2016). Perspectives on genetically modified crops and food detection. Journal of Food and Drug Analysis 24(1): 1-8.
28. Lipp M, Brodmann P, Pietsch K, Pauwels J, Anklam E (1999). IUPAC collaborative trial study of a method to detect genetically modified soy beans and maize in dried powder. Journal of AOAC International 82(4): 923-928.
29. Lipp M, Bluth A, Eyquem F, Kruse L, Schimmel H, Van den Eede G, Anklam E (2001). Validation of a method based on polymerase chain reaction for the detection of genetically modified organisms in various processed foodstuffs. European Food Research and Technology 212: 497-504.
30. Mafra I, Silva SA, Moreira EJ, da Silva CSF, Beatriz M, Oliveira PP (2008). Comparative study of DNA extraction methods for soybean derived food products. Food Control 19(12): 1183-1190.
31. Mutlu Ş, Şimşek O, Öksüz Ö (2021). Investigating The GMO Existence in Chips and Breakfast Cereals Marketed in Turkey. Tekirdağ Ziraat Fakültesi Dergisi 18(3): 375-385.
32. Ohmori K, Hisayo T, Takahiro W, Hiroshi A, Tamio M, Toshiharu Y, Shin-Ichi I, Shuji S (2008). Method for DNA Extraction and Purification from Corn-processed Food Using an Ion-exchange Resin Type Kit. J-Stage 49(1): 45-50.
33. Pacheco Coello R, Pestana Justo J, Factos Mendoza A, Santos Ordoñez E (2017). Comparison of three DNA extraction methods for the detection and quantification of GMO in Ecuadorian manufactured food. BMC Research Notes 10: 1-7.
34. Peano C, Samson MC, Palmieri L, Gulli M, Marmiroli N (2004). Qualitative and quantitative evaluation of the genomic DNA extracted from GMO and non-GMO foodstuffs with four different extraction methods. Journal of agricultural and food chemistry 52(23): 6962-6968.
35. Piskata Z, Servusova E, Babak V, Nesvadbova M, Borilova G (2019). The quality of DNA isolated from processed food and feed via different extraction procedures. Molecules 24(6): 1188.
36. Pirondini A, Bonas U, Maestri E, Visioli G, Marmiroli M, Marmiroli N (2010). Yield and amplificability of different DNA extraction procedures for traceability in the dairy food chain. Food Control 21: 663–668.
37. Ramos-Gómez S, Busto MD, Perez-Mateos M, Ortega N (2014). Development of a method to recovery and amplification DNA by real-time PCR from commercial vegetable oils. Food chemistry 158: 374-383.
38. Sajali N, Wong SC, Hanapi UK, Abu Bakar@ Jamaluddin, S, Tasrip NA, Mohd Desa MN (2018). The challenges of DNA extraction in different assorted food matrices: a review. Journal of Food Science 83(10): 2409-2414.
39. Sahu SK, Thangaraj M, Kathiresan K (2012). DNA extraction protocol for plants with high levels of secondary metabolites and polysaccharides without using liquid nitrogen and phenol. International Scholarly Research Notices 2012(1): 205049.
40. Sambrook J, Russell D (2001). Molecular cloning: a laboratory manual, 3rd ed. New York: Cold Spring Harbor Laboratory Press.
41. Singh M, Sodhi KK, Paliwal A, Sharma S, Randhawa G (2021). Efficient DNA extraction procedures for processed food derivatives - a critical step to ensure quality for GMO analysis. Food Analytical Methods 14(11): 2249-2261.
42. Sisea C, Pamfil D (2007). Comparison of DNA extraction methods for GMO analysis of food products. Bull USAMV-CN 63, 64.
43. Ateş Sönmezoglu Ö, Keskin H (2015). Determination of genetically modified corn and soy in processed food products. Journal of Applied Biology and Biotechnology 3(3): 32-37.
44. Soylu BB, Erkan I, Yukseloğlu EH (2020). Searching of the genetically modified organisms and their products’ status and evaluation of food safety and regulations in Turkey in terms of the Forensic Sciences. Commagene Journal of Biology 4(2): 104-109.
45. Tan SC, Yiap BC (2009). DNA, RNA, and protein extraction: the past and the present. (2009: 574398. Published online 2009 November 30). J Biomed Biotechnol.
46. Turkec A, Kazan H, Karacanli B, Lucas SJ (2015). DNA extraction techniques compared for accurate detection of genetically modified organisms (GMOs) in maize food and feed products. Journal of Food Science and Technology 52: 5164-5171.
47. Tung-Nguyen CT, Son R, Raha AR, Lai OM, Clemente Michael WVL (2009). Comparison of DNA extraction efficiencies using various methods for the detection of genetically modified organisms (GMOs). Int Food Research Journal 16: 21-30.
48. Vahdani M, Sahari MA, Tanavar M (2024). Quantitative and qualitative analysis of three DNA extraction methods from soybean, maize, and canola oils and investigation of the presence of genetically modified organisms (GMOs). Food Chemistry: Molecular Sciences 8: 100201.
49. Verginelli D, Spinella K, La Rocca D, Bonini P, Fusco C, Misto M, Marchesi U (2023). Validation and comparison of four DNA extraction methods for genetically modified organisms analysis: From DNA quality control to quantification of genetically modified content in real‐life samples matrices. JSFA Reports 3(11): 519-535.
50. Vijayakumar KR, Martin A, Gowda LR, Prakash V (2009). Detection of genetically modified soya and maize: Impact of heat processing. Food Chemistry 117(3): 514-521.
51. Walker JM, Wilson K (Eds.) (2010). Principles and Techniques of biochemistry and molecular biology. Cambridge: Cambridge University Press.
52. Wang X, Teng D, Tian F, Guan Q, Wang J (2012). Comparison of three DNA extraction methods for feed products and four amplification methods for the 5′-junction fragment of roundup ready soybean. Journal of Agricultural and Food Chemistry 60(18): 4586-4595.
53. Wilfinger WW, Mackey K, Chomczynski P (2006). Assessing the Quantity; Purity and Integrity of RNA and DNA following Nucleic acid purification. DNA Sequencing II Optimising Preparation and Cleanup 291-312.
54. Yönetmelik (TR) (2010). Genetiği değiştirilmiş organizmalar ve ürünlerine ilişkin Türk mevzuatı; Resmi Gazete No: 27671. 2010. http://www.resmigazete.gov.tr/eskiler/2010/08/20100813-4.htm [accessed 13.12.2023].
55. Xia Y, Chen F, Du Y, Liu C, Bu G, Xin Y, Liu B (2019). A modified SDS-based DNA extraction method from raw soybean. Bioscience Reports 39(2): BSR20182271.