A multiplex degenerate PCR analytical approach targeting to eight genes for screening GMOs

• Published in: Food chemistry Vol. 132; no. 3; pp. 1566 - 1573
• Main Authors: Guo, Jinchao, Chen, Lili, Liu, Xin, Gao, Ying, Zhang, Dabing, Yang, Litao
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier Ltd 01.06.2012; Elsevier
• Subjects: Biological and medical sciences; Food industries; Fundamental and applied biological sciences. Psychology; Genetically modified organisms; Quadruplex degenerate PCR; Trait gene; Quadruplex degenerate PCR; Genetically modified organisms; Trait gene; Polymerase chain reaction; Screening; Analysis method; Targeting; Gene; Molecular biology; Genetically modified organism
• ISSN: 0308-8146; 1873-7072
• DOI: 10.1016/j.foodchem.2011.11.096

► One quadruplex PCR assay for screening the presence of GMO targets in foods/feeds. ► This quadruplex PCR assay comprises four degenerate primer pairs targeted on eight exogenous DNA targets in GMOs. ► This developed assay could be used for screening more than 90 authorized GM events. ► The developed assay has been successfully used for screening the artificial GM samples and GIPSA proficiency testing samples. Currently, the detection methods with lower cost and higher throughput are the major trend in screening genetically modified (GM) food or feed before specific identification. In this study, we developed a quadruplex degenerate PCR screening approach for more than 90 approved GMO events. This assay is consisted of four PCR systems targeting on nine DNA sequences from eight trait genes widely introduced into GMOs, such as CP4-EPSPS derived from Acetobacterium tumefaciens sp. strain CP4, phosphinothricin acetyltransferase gene derived from Streptomyceshygroscopicus (bar) and Streptomyces viridochromogenes (pat), and Cry1Ab, Cry1Ac, Cry1A(b/c), mCry3A, and Cry3Bb1 derived from Bacillus thuringiensis. The quadruplex degenerate PCR assay offers high specificity and sensitivity with the absolute limit of detection (LOD) of approximate 80targetcopies. Furthermore, the applicability of the quadruplex PCR assay was confirmed by screening either several artificially prepared samples or samples of Grain Inspection, Packers and Stockyards Administration (GIPSA) proficiency program.