Simultaneous Detection of Eight Genetically Modified Maize Lines Using a Combination of Event- and Construct-Specific Multiplex-PCR Technique

• Published in: Journal of agricultural and food chemistry Vol. 56; no. 19; pp. 8962 - 8968
• Main Authors: Shrestha, Hari K, Hwu, Kae-Kang, Wang, Shu-Jen, Liu, Li-Fei, Chang, Men-Chi
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 08.10.2008
• Subjects: Biological and medical sciences; Cereal and baking product industries; Chemical Aspects of Biotechnology/Molecular Biology; construct-specific; corn; detection; DNA, Plant - analysis; DNA, Plant - chemistry; event-specific; food analysis; Food industries; Fundamental and applied biological sciences. Psychology; General aspects; genes; Genetically modified (GM) maize (Zea maysL.); genetically modified foods; Methods of analysis, processing and quality control, regulation, standards; multiplex-PCR (mPCR); Plants, Genetically Modified - genetics; polymerase chain reaction; Polymerase Chain Reaction - methods; screening; Sequence Analysis, DNA; transgene integration sequence; transgenic plants; Zea mays; Zea mays - genetics; zein; multiplex-PCR (mPCR); Genetically modified (GM) maize (Zea mays L.); event-specific; construct-specific; transgene integration sequence; Monocotyledones; Zea mays; multiplex- PCR (mPCR); Corn; Polymerase chain reaction; Analysis method; Gramineae; Angiospermae; Cereal; Spermatophyta; Control method; Molecular biology; Detection; Genetically modified organism
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/jf800501z

To fulfill labeling and traceability requirement of genetically modified (GM) maize for trade and regulation, it is essential to develop an event-specific detection method for monitoring the presence of transgenes. In pursuit of this purpose, we systematically optimized and established a combined event- and construct-specific multiplex polymerase chain reaction (mPCR) technique for simultaneous detection of 8 GM maize lines. Altogether 9 sets of primers were designed, including six that were event-specific for Event176, Bt11, TC1507, NK603, MON863, and Mon810; two that were construct-specific for T25 and GA21, and one for an endogenous zein gene. The transgene in each GM maize line and the endogenous zein gene could be clearly detected and distinguished according to the different sizes of PCR amplicons. The limit of detection (LOD) was approximately 0.25% (v/v), although the detection can be as sensitive as 0.1% as demonstrated by the International Seed Testing Association (ISTA) proficiency test. This study further improves the current PCR-based detection method for GM maize. The method can be used in an easy, sensitive, and cost and time effective way for the identification and quality screening of a specific GM maize line.