title: Assessment of DNA degradation induced by thermal and UV radiation processing: implications for quantification of genetically modified organisms.
abstract: DNA quality is an important parameter for the detection and quantification of genetically modified organisms (GMO's) using the polymerase chain reaction (PCR). Food processing leads to degradation of DNA, which may impair GMO detection and quantification. This study evaluated the effect of various processing treatments such as heating, baking, microwaving, autoclaving and ultraviolet (UV) irradiation on the relative transgenic content of MON 810 maize using pRSETMON-02, a dual target plasmid as a model system. Amongst all the processing treatments examined, autoclaving and UV irradiation resulted in the least recovery of the transgenic (CaMV 35S promoter) and taxon-specific (zein) target DNA sequences. Although a profound impact on DNA degradation was seen during the processing, DNA could still be reliably quantified by Real-time PCR. The measured mean DNA copy number ratios of the processed samples were in agreement with the expected values. Our study confirms the premise that the final analytical value assigned to a particular sample is independent of the degree of DNA degradation since the transgenic and the taxon-specific target sequences possessing approximately similar lengths degrade in parallel. The results of our study demonstrate that food processing does not alter the relative quantification of the transgenic content provided the quantitative assays target shorter amplicons and the difference in the amplicon size between the transgenic and taxon-specific genes is minimal.
authors.affiliation: Department of Food Safety and Analytical Quality Control Laboratory, CSIR-Central Food Technological Research Institute, Mysore 570 020, India.
authors.forename: Rajashekhar V
authors.initials: RV
authors.lastname: Ballari
authors.forename: Asha
authors.initials: A
authors.lastname: Martin
mesh_terms: D018744:DNA, Plant / Q000235:genetics*; D005511:Food Handling; D005514:Food Irradiation; D006358:Hot Temperature; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000528:radiation effects*; D010957:Plasmids / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D014466:Ultraviolet Rays; D003313:Zea mays / Q000235:genetics* / Q000528:radiation effects*; D015028:Zein / Q000235:genetics
chemical_list: D018744:DNA, Plant; D015028:Zein
keywords: DNA degradation; Food processing; GMO quantification; Genetically modified organisms; Real-time PCR; UV irradiation